Bicyclic inhibitors of Rho kinase

ABSTRACT

The present invention relates to compounds useful as inhibitors of protein kinases, particularly of ROCK. The invention also provides pharmaceutically acceptable compositions comprising said compounds and methods of using the compositions in the treatment of various disease, conditions, or disorders.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/630,115, filed Nov. 22, 2005, which is herein incorporated byreference.

TECHNICAL FIELD OF INVENTION

The present invention relates to compounds useful as inhibitors ofprotein kinases. The invention also provides pharmaceutically acceptablecompositions comprising the compounds of the invention and methods ofusing the compositions in the treatment of various disorders.

BACKGROUND OF THE INVENTION

Protein kinases constitute a large family of structurally relatedenzymes that are responsible for the control of a variety of signaltransduction processes within the cell. Many diseases are associatedwith abnormal cellular responses triggered by protein kinase-mediatedevents. Accordingly, there has been a substantial effort in medicinalchemistry to find protein kinase inhibitors that are effective astherapeutic agents.

One kinase family of interest is Rho-associated coiled-coil formingprotein serine/threonine kinase (ROCK), which is believed to be aneffector of Ras-related small GTPase Rho. The ROCK family includesp160ROCK (ROCK-1) and ROKα/Rho-kinase/ROCK-II, protein kinase PKN, andcitron and citron kinase. The ROCK family of kinases have been shown tobe involved in a variety of functions including Rho-induced formation ofactin stress fibers and focal adhesions and in downregulation of myosinphosphatase, platelet activation, aortic smooth muscle contraction byvarious stimuli, thrombin-induced responses of aortic smooth musclecells, hypertrophy of cardiomyocytes, bronchial smooth musclecontraction, smooth muscle contraction and cytoskeletal reorganizationof non-muscle cells, activation of volume-regulated anion channels,neurite retraction, neutrophil chemotaxis, wound healing, tumor invasionand cell transformation. More specifically, ROCK has been implicated invarious diseases and disorders including hypertension, cerebralvasospasm, coronary vasospasm, bronchial asthma, preterm labor, erectiledysfunction, glaucoma, vascular smooth muscle cell proliferation,myocardial hypertrophy, malignoma, ischemia/reperfusion-induced injury,endothelial dysfunction, Crohn's Disease and colitis, neurite outgrowth,Raynaud's Disease, angina, Alzheimer's disease, benign prostatichyperplasia and atherosclerosis. Accordingly, the development ofinhibitors of ROCK kinase would be useful as therapeutic agents for thetreatment of disorders implicated in the ROCK kinase pathway.

Accordingly, there is a great need to develop inhibitors of ROCK thatwould be useful in treating various diseases or conditions associatedwith ROCK, particularly given the inadequate treatments currentlyavailable for the majority of these disorders.

SUMMARY OF THE INVENTION

It has now been found that compounds of this invention, andpharmaceutically acceptable compositions thereof, are effective asinhibitors of ROCK. These compounds have the general formula I:

or a pharmaceutically acceptable derivative thereof, wherein ring B, Z¹,Z², Z³, Z⁴, R³, G and Q¹ are as defined below.

These compounds, and pharmaceutically acceptable compositions thereof,are useful for treating or lessening the severity of a variety ofdisorders, including, without limitation, hypertension, cerebralvasospasm, coronary vasospasm, bronchial asthma, preterm labor, erectiledysfunction, glaucoma, vascular smooth muscle cell proliferation,myocardial hypertrophy, malignoma, ischemia/reperfusion-induced injury,endothelial dysfunction, Crohn's Disease and colitis, neurite outgrowth,Raynaud's Disease, angina, Alzheimer's disease, benign prostatichyperplasia and atherosclerosis.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

As used herein, the following definitions shall apply unless otherwiseindicated. For purposes of this invention, the chemical elements areidentified in accordance with the Periodic Table of the Elements, CASversion, and the Handbook of Chemistry and Physics, 75^(th) Ed. 1994.Additionally, general principles of organic chemistry are described in“Organic Chemistry”, Thomas Sorrell, University Science Books,Sausalito: 1999, and “March's Advanced Organic Chemistry”, 5^(th) Ed.,Smith, M. B. and March, J., eds. John Wiley & Sons, New York: 2001, theentire contents of which are hereby incorporated by reference.

As described herein, compounds of the invention may optionally besubstituted with one or more substituents, such as are illustratedgenerally above, or as exemplified by particular classes, subclasses,and species of the invention. It will be appreciated that the phrase“optionally substituted” is used interchangeably with the phrase“substituted or unsubstituted.” In general, the term “substituted”,whether preceded by the term “optionally” or not, refers to thereplacement of one or more hydrogen radicals in a given structure withthe radical of a specified substituent. Unless otherwise indicated, anoptionally substituted group may have a substituent at eachsubstitutable position of the group. When more than one position in agiven structure can be substituted with more than one substituentselected from a specified group, the substituent may be either the sameor different at each position.

As described herein, when the term “optionally substituted” precedes alist, said term refers to all of the subsequent substitutable groups inthat list. If a substituent radical or structure is not identified ordefined as “optionally substituted”, the substituent radical orstructure is unsubstituted. For example, if X is halogen; optionallysubstituted C₁₋₃alkyl or phenyl; X may be either optionally substitutedalkyl or optionally substituted phenyl. Likewise, if the term“optionally substituted” follows a list, said term also refers to all ofthe substitutable groups in the prior list unless otherwise indicated.For example: if X is halogen, C₁₋₃alkyl or phenyl wherein X isoptionally substituted by J^(X), then both C₁₋₃alkyl and phenyl may beoptionally substituted by J^(X). As is apparent to one having ordinaryskill in the art, groups such as H, halogen, NO₂, CN, NH₂, OH, or OCF₃would not be included because they are not substitutable groups.

Combinations of substituents envisioned by this invention are preferablythose that result in the formation of stable or chemically feasiblecompounds. The term “stable”, as used herein, refers to compounds thatare not substantially altered when subjected to conditions to allow fortheir production, detection, and, preferably, their recovery,purification, and use for one or more of the purposes disclosed herein.In some embodiments, a stable compound or chemically feasible compoundis one that is not substantially altered when kept at a temperature of40° C. or less, in the absence of moisture or other chemically reactiveconditions, for at least a week.

The term “aliphatic” or “aliphatic group”, as used herein, means astraight-chain (i.e., unbranched) or branched, substituted orunsubstituted hydrocarbon chain that is completely saturated or thatcontains one or more units of unsaturation. Unless otherwise specified,aliphatic groups contain 1-20 aliphatic carbon atoms. In someembodiments, aliphatic groups contain 1-10 aliphatic carbon atoms. Inother embodiments, aliphatic groups contain 1-8 aliphatic carbon atoms.In still other embodiments, aliphatic groups contain 1-6 aliphaticcarbon atoms, and In yet other embodiments, aliphatic groups contain 1-4aliphatic carbon atoms. Suitable aliphatic groups include, but are notlimited to, linear or branched, substituted or unsubstituted alkyl,alkenyl, or alkynyl groups. Further examples of aliphatic groups includemethyl, ethyl, propyl, butyl, isopropyl, isobutyl, vinyl, and sec-butyl.

The term “cycloaliphatic” (or “carbocycle” or “cycloalkyl”) refers to amonocyclic C₃-C₈ hydrocarbon or bicyclic C₈-C₁₂ hydrocarbon that iscompletely saturated or that contains one or more units of unsaturation,but which is not aromatic, that has a single point of attachment to therest of the molecule, and wherein any individual ring in said bicyclicring system has 3-7 members. Suitable cycloaliphatic groups include, butare not limited to, cycloalkyl, cycloalkenyl, and cycloalkynyl. Furtherexamples of aliphatic groups include cyclopentyl, cyclopentenyl,cyclohexyl, cyclohexenyl, cycloheptyl, and cycloheptenyl.

The term “heterocycle”, “heterocyclyl”, “heterocycloaliphatic”, or“heterocyclic” as used herein refers to a monocyclic, bicyclic, ortricyclic ring system in which one or more ring members are anindependently selected heteroatom and that is completely saturated orthat contains one or more units of unsaturation, but which is notaromatic, that has a single point of attachment to the rest of themolecule. In some embodiments, the “heterocycle”, “heterocyclyl”,“heterocycloaliphatic”, or “heterocyclic” group has three to fourteenring members in which one or more ring members is a heteroatomindependently selected from oxygen, sulfur, nitrogen, or phosphorus, andeach ring in the system contains 3 to 7 ring members.

Examples of heterocyclic rings include, but are not limited to, thefollowing monocycles: 2-tetrahydrofuranyl, 3-tetrahydrofuranyl,2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl, 2-morpholino,3-morpholino, 4-morpholino, 2-thiomorpholino, 3-thiomorpholino,4-thiomorpholino, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl,1-tetrahydropiperazinyl, 2-tetrahydropiperazinyl,3-tetrahydropiperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl,1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl,1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl,2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl,2-imidazolidinyl, 4-imidazolidinyl, 5-imidazolidinyl; and the followingbicycles: 3-1H-benzimidazol-2-one, 3-(1-alkyl)-benzimidazol-2-one,indolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane,benzodithiane, and 1,3-dihydro-imidazol-2-one.

The term “heteroatom” means one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon, including any oxidized form of nitrogen, sulfur,phosphorus, or silicon, the quaternized form of any basic nitrogen, or asubstitutable nitrogen of a heterocyclic ring, for example N (as in3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR⁺ (as inN-substituted pyrrolidinyl).

The term “unsaturated”, as used herein, means that a moiety has one ormore units of unsaturation.

The term “alkoxy”, or “thioalkyl”, as used herein, refers to an alkylgroup, as previously defined, attached to the principal carbon chainthrough an oxygen (“alkoxy”) or sulfur (“thioalkyl”) atom.

The terms “haloalkyl”, “haloalkenyl” and “haloalkoxy” means alkyl,alkenyl or alkoxy, as the case may be, substituted with one or morehalogen atoms. The term “halogen” means F, Cl, Br, or I.

The term “aryl” used alone or as part of a larger moiety as in“aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refers to monocyclic,bicyclic, and tricyclic carbocyclic ring systems having a total of sixto fourteen ring members, wherein at least one ring in the system isaromatic, wherein each ring in the system contains 3 to 7 ring membersand that has a single point of attachment to the rest of the molecule.The term “aryl” may be used interchangeably with the term “aryl ring”.Examples of aryl rings would include phenyl, naphthyl, and anthracene.

The term “heteroaryl”, used alone or as part of a larger moiety as in“heteroaralkyl” or “heteroarylalkoxy”, refers to monocyclic, bicyclic,and tricyclic ring systems having a total of five to fourteen ringmembers, wherein at least one ring in the system is aromatic, at leastone ring in the system contains one or more heteroatoms, wherein eachring in the system contains 3 to 7 ring members and that has a singlepoint of attachment to the rest of the molecule. The term “heteroaryl”may be used interchangeably with the term “heteroaryl ring” or the term“heteroaromatic”.

Further examples of heteroaryl rings include the following monocycles:2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl,5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl,4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl,3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl,pyridazinyl (e.g., 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl,5-thiazolyl, tetrazolyl (e.g., 5-tetrazolyl), triazolyl (e.g.,2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, pyrazolyl (e.g.,2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl,1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl,1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, pyrazinyl, 1,3,5-triazinyl, andthe following bicycles: benzimidazolyl, benzofuryl, benzothiophenyl,indolyl (e.g., 2-indolyl), purinyl, quinolinyl (e.g., 2-quinolinyl,3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl,3-isoquinolinyl, or 4-isoquinolinyl).

In some embodiments, an aryl (including aralkyl, aralkoxy, aryloxyalkyland the like) or heteroaryl (including heteroaralkyl andheteroarylalkoxy and the like) group may contain one or moresubstituents. Suitable substituents on the unsaturated carbon atom of anaryl or heteroaryl group are selected from those listed in thedefinition of J^(Q), J^(R), J^(V), J^(U) and J^(X) below. Other suitablesubstituents include: halogen; —R^(o); —OR^(o); —SR^(o);1,2-methylenedioxy; 1,2-ethylenedioxy; phenyl (Ph) optionallysubstituted with R^(o); —O(Ph) optionally substituted with R^(o);—(CH₂)₁₋₂(Ph), optionally substituted with R^(o); —CH═CH(Ph), optionallysubstituted with R^(o); —NO₂; —CN; —N(R^(o))₂; —NR^(o)C(O)R^(o);—NR^(o)C(S)R^(o); —NR^(o)C(O)N(R^(o))₂; —NR^(o)C(S)N(R^(o))₂;—NR^(o)CO₂R^(o); —NR^(o)NR^(o)C(O)R^(o); —NR^(o)NR^(o)C(O)N(R^(o))₂;—NR^(o)NR^(o)CO₂R^(o); —C(O)C(O)R^(o); —C(O)CH₂C(O)R^(o); —CO₂R^(o);—C(O)R^(o); —C(S)R^(o); —C(O)N(R^(o))₂; —C(S)N(R^(o))₂; —OC(O)N(R^(o))₂;—OC(O)R^(o); —C(O)N(OR^(o))R^(o); —C(NOR^(o))R^(o); —S(O)₂R^(o);—S(O)₃R^(o); —SO₂N(R^(o))₂; —S(O)R^(o); —NR^(o)SO₂N(R^(o))₂;—NR^(o)SO₂R^(o); —N(OR^(o))R^(o); —C(═NH)—N(R^(o))₂; or—(CH₂)₀₋₂NHC(O)R^(o); wherein each independent occurrence of R^(o) isselected from hydrogen, optionally substituted C₁₋₆ aliphatic, anunsubstituted 5-6 membered heteroaryl or heterocyclic ring, phenyl,—O(Ph), or —CH₂(Ph), or, two independent occurrences of R^(o), on thesame substituent or different substituents, taken together with theatom(s) to which each R^(o) group is bound, form a 5-8-memberedheterocyclyl, aryl, or heteroaryl ring or a 3-8-membered cycloalkylring, wherein said heteroaryl or heterocyclyl ring has 1-3 heteroatomsindependently selected from nitrogen, oxygen, or sulfur. Optionalsubstituents on the aliphatic group of R^(o) are selected from NH₂,NH(C₁₋₄aliphatic), N(C₁₋₄aliphatic)₂, halogen, C₁₋₄aliphatic, OH,O(C₁₋₄aliphatic), NO₂, CN, CO₂H, CO₂(C₁₋₄aliphatic), O(haloC₁₋₄aliphatic), or haloC₁₋₄aliphatic, wherein each of the foregoingC₁₋₄aliphatic groups of R^(o) is unsubstituted.

In some embodiments, an aliphatic or heteroaliphatic group, or anon-aromatic heterocyclic ring may contain one or more substituents.Suitable substituents on the saturated carbon of an aliphatic orheteroaliphatic group, or of a non-aromatic heterocyclic ring areselected from those listed above for the unsaturated carbon of an arylor heteroaryl group and additionally include the following: ═O, ═S,═NNHR*, ═NN(R*)₂, ═NNHC(O)R*, ═NNHCO₂(alkyl), ═NNHSO₂(alkyl), or ═NR*,where each R* is independently selected from hydrogen or an optionallysubstituted C₁₋₆ aliphatic. Optional substituents on the aliphatic groupof R* are selected from NH₂, NH(C₁₋₄ aliphatic), N(C₁₋₄ aliphatic)₂,halogen, C₁₋₄ aliphatic, OH, O(C₁₋₄ aliphatic), NO₂, CN, CO₂H, CO₂(C₁₋₄aliphatic), O(halo C₁₋₄ aliphatic), or halo(C₁₋₄ aliphatic), whereineach of the foregoing C₁₋₄aliphatic groups of R* is unsubstituted.

In some embodiments, optional substituents on the nitrogen of anon-aromatic heterocyclic ring include —R⁺, —N(R⁺)₂, —C(O)R⁺, —CO₂R⁺,—C(O)C(O)R⁺, —C(O)CH₂C(O)R⁺, —SO₂R⁺, —SO₂N(R⁺)₂, —C(═S)N(R⁺)₂,—C(═NH)—N(R⁺)₂, or —NR⁺SO₂R⁺; wherein R⁺ is hydrogen, an optionallysubstituted C₁₋₆ aliphatic, optionally substituted phenyl, optionallysubstituted —O(Ph), optionally substituted —CH₂(Ph), optionallysubstituted —(CH₂)₁₋₂(Ph); optionally substituted —CH═CH(Ph); or anunsubstituted 5-6 membered heteroaryl or heterocyclic ring having one tofour heteroatoms independently selected from oxygen, nitrogen, orsulfur, or, two independent occurrences of R⁺, on the same substituentor different substituents, taken together with the atom(s) to which eachR⁺ group is bound, form a 5-8-membered heterocyclyl, aryl, or heteroarylring or a 3-8-membered cycloalkyl ring, wherein said heteroaryl orheterocyclyl ring has 1-3 heteroatoms independently selected fromnitrogen, oxygen, or sulfur. Optional substituents on the aliphaticgroup or the phenyl ring of R⁺ are selected from NH₂, NH(C₁₋₄aliphatic), N(C₁₋₄ aliphatic)₂, halogen, C₁₋₄ aliphatic, OH, O(C₁₋₄aliphatic), NO₂, CN, CO₂H, CO₂(C₁₋₄ aliphatic), O(halo C₁₋₄ aliphatic),or halo(C₁₋₄ aliphatic), wherein each of the foregoing C₁₋₄aliphaticgroups of R⁺ is unsubstituted.

As detailed above, in some embodiments, two independent occurrences ofR^(o) (or R⁺, or any other variable similarly defined herein), may betaken together with the atom(s) to which each variable is bound to forma 5-8-membered heterocyclyl, aryl, or heteroaryl ring or a 3-8-memberedcycloalkyl ring. Exemplary rings that are formed when two independentoccurrences of R^(o) (or R⁺, or any other variable similarly definedherein) are taken together with the atom(s) to which each variable isbound include, but are not limited to the following: a) two independentoccurrences of R^(o) (or R⁺, or any other variable similarly definedherein) that are bound to the same atom and are taken together with thatatom to form a ring, for example, N(R^(o))₂, where both occurrences ofR^(o) are taken together with the nitrogen atom to form apiperidin-1-yl, piperazin-1-yl, or morpholin-4-yl group; and b) twoindependent occurrences of R^(o) (or R⁺, or any other variable similarlydefined herein) that are bound to different atoms and are taken togetherwith both of those atoms to form a ring, for example where a phenylgroup is substituted with two occurrences of OR^(o)

these two occurrences of R^(o) are taken together with the oxygen atomsto which they are bound to form a fused 6-membered oxygen containingring:

It will be appreciated that a variety of other rings can be formed whentwo independent occurrences of R^(o) (or R⁺, or any other variablesimilarly defined herein) are taken together with the atom(s) to whicheach variable is bound and that the examples detailed above are notintended to be limiting.

In some embodiments, an alkyl or aliphatic chain can be optionallyinterrupted with another atom or group. This means that a methylene unitof the alkyl or aliphatic chain is optionally replaced with said otheratom or group. Examples of such atoms or groups would include, but arenot limited to, —NR—, —O—, —S—, —CO₂—, —OC(O)—, —C(O)CO—, —C(O)—,—C(O)NR—, —C(═N—CN), —NRCO—, —NRC(O)O—, —SO₂NR—, —NRSO₂—, —NRC(O)NR—,—OC(O)NR—, —NRSO₂NR—, —SO—, or —SO₂—, wherein R is defined herein.Unless otherwise specified, the optional replacements form a chemicallystable compound. Optional interruptions can occur both within the chainand at either end of the chain; i.e. both at the point of attachmentand/or also at the terminal end. Two optional replacements can also beadjacent to each other within a chain so long as it results in achemically stable compound. Unless otherwise specified, if thereplacement or interruption occurs at the terminal end, the replacementatom is bound to an H on the terminal end. For example, if —CH₂CH₂CH₃were optionally interrupted with —O—, the resulting compound could be—OCH₂CH₃, —CH₂OCH₃, or —CH₂CH₂OH.

As described herein, a bond drawn from a substituent to the center ofone ring within a multiple-ring system (as shown below), representssubstitution of the substituent at any substitutable position in any ofthe rings within the multiple ring system. For example, Figure arepresents possible substitution in any of the positions shown in Figureb.

This also applies to multiple ring systems fused to optional ringsystems (which would be represented by dotted lines). For example, inFigure c, X is an optional substituent both for ring A and ring B.

If, however, two rings in a multiple ring system each have differentsubstituents drawn from the center of each ring, then, unless otherwisespecified, each substituent only represents substitution on the ring towhich it is attached. For example, in Figure d, Y is an optionallysubstituent for ring A only, and X is an optional substituent for ring Bonly.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, (Z) and (E) double bondisomers, and (Z) and (E) conformational isomers. Therefore, singlestereochemical isomers as well as enantiomeric, diastereomeric, andgeometric (or conformational) mixtures of the present compounds arewithin the scope of the invention.

Unless otherwise stated, all tautomeric forms of the compounds of theinvention are within the scope of the invention. Additionally, unlessotherwise stated, structures depicted herein are also meant to includecompounds that differ only in the presence of one or more isotopicallyenriched atoms. For example, compounds having the present structuresexcept for the replacement of hydrogen by deuterium or tritium, or thereplacement of a carbon by a ¹³C- or ¹⁴C-enriched carbon are within thescope of this invention. Such compounds are useful, for example, asanalytical tools or probes in biological assays.

As represented herein, the left-hand bond of Ring B is attached to thebicyclic heteroaryl ring and the right-hand bond of Ring B is attachedto radical G.

Description of Compounds of the Invention:

The present invention relates to a compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein:wherein

-   Z₁, Z₂, Z₃ and Z₄ are each independently selected from N or CR¹,    wherein at least one of Z₁, Z₂, or Z₄ is N;-   each R¹ is independently selected from H, halogen, —CN, —NO₂, or    —V_(m)R′;-   G is —NR²— or —CO—;-   Q¹ is —CO—, —SO₂—, —NR², —NR²CO—, —CONR²—, —SO₂NR²—, or is a bond;-   R² is —U_(n)R″;-   R³ is Q²-Ar¹, or when G is —NR², R² and Q¹-R³, taken together with    the nitrogen atom, may form the cyclic group:    where s is 1 or 2, Z is CH or N; wherein each occurrence of Y is    independently —CO—, —CS—, —SO₂—, —O—, —S—, —NR⁵—, or —C(R⁵)₂—, and    R⁵ is U_(n)R′;-   X₁ and X₂ are each independently selected from CR⁴ or N;-   each occurrence of R⁴ is independently selected from halogen, CN,    NO₂, or V_(m)R;-   each occurrence of U or V is independently selected from an    optionally substituted C₁₋₆ alkylidene chain, wherein up to two    methylene units of the chain are optionally and independently    replaced by —NR—, —S—, —O—, —CS—, —CO₂—, —OCO—, —CO—, —COCO—,    —CONR—, —NRCO—, —NRCO₂—, —SO₂NR—, —NRSO₂—, —CONRNR—, —NRCONR—,    —OCONR—, —NRNR—, —NRSO₂NR—, —SO—, —SO₂—, —PO—, —PO₂—, or —POR—;-   m and n are each independently 0 or 1;-   each occurrence of R is independently selected from hydrogen or a    C₁₋₆ aliphatic group, wherein said aliphatic group is optionally    substituted with up to five occurrences of J^(R);-   each occurrence of R′ is independently selected from hydrogen, a    C₁₋₆ aliphatic group, a 3-8-membered saturated, partially    unsaturated, or fully unsaturated monocyclic ring having 0-3    heteroatoms independently selected from nitrogen, oxygen, or sulfur,    or an 8-12 membered saturated, partially unsaturated, or fully    unsaturated bicyclic ring system having 0-5 heteroatoms    independently selected from nitrogen, oxygen, or sulfur, wherein    said aliphatic group, monocyclic ring or bicyclic ring is optionally    substituted with up to five occurrences of J^(R′);-   R″ is selected from hydrogen, a C₁₋₆ aliphatic group, a 3-8-membered    saturated, partially unsaturated, or fully unsaturated monocyclic    ring having 0-3 heteroatoms independently selected from nitrogen,    oxygen, or sulfur, or an 8-12 membered saturated, partially    unsaturated, or fully unsaturated bicyclic ring system having 0-5    heteroatoms independently selected from nitrogen, oxygen, or sulfur,    wherein said aliphatic group, monocyclic ring or bicyclic ring is    optionally substituted with up to five occurrences of J^(R″);-   or two occurrences of R, R′ and R″, in any combination thereof, are    taken together with the atom(s) to which they are bound to form a    3-12 membered saturated, partially unsaturated, or fully unsaturated    monocyclic or bicyclic ring having 0-4 heteroatoms independently    selected from nitrogen, oxygen, or sulfur, wherein said monocyclic    or bicyclic ring is optionally substituted with J^(R);-   each occurrence of J^(R), J^(R′) and J^(R″) is independently    selected from halogen, L, -(L_(p))-R^(J), -(L_(p))-N(R^(J))₂,    -(L_(p))-SR^(J), -(L_(p))-OR^(J), -(L_(p))-(C₃₋₁₀ cycloaliphatic),    -(L_(p))-(C₆₋₁₀ aryl), -(L_(p))-(5-10 membered heteroaryl),    -(L_(p))-(5-10 membered heterocyclyl), oxo, C₁₋₄haloalkoxy,    C₁₋₄haloalkyl, -(L_(p))-NO₂, -(L_(p))-CN, -(L_(p))-OH, -(L_(p))-CF₃,    —CO₂R^(J), —CO₂H, —COR^(J), —COH, —OC(O)R^(J) or —NC(O)R^(J); or any    two J^(R), J^(R′) or J^(R″) groups, on the same substituent or    different substituents, together with the atom(s) to which each    J^(R), J^(R′) or J^(R″) group is bound, form a 5-7 membered    saturated, unsaturated, or partially saturated ring;-   R^(J) is H or C₁₋₆ aliphatic; or two R^(J) groups or an R^(J) group    and an R, R′ or R″ group, together with the atom to which they are    attached, optionally form a 3-6 membered cycloaliphatic or    heterocyclyl, wherein said aliphatic, cycloaliphatic or heterocyclyl    is optionally substituted with R*, —OR*, —SR*, —NO₂, —CF₃, —CN,    —CO₂R*, —COR*, OCOR* or NHCOR*, wherein R* is H or an unsubstituted    C₁₋₆ aliphatic;-   L is a C₁₋₆ aliphatic wherein up to three methylene units are    replaced by —NH—, —NR^(L), —O—, —S—, —CO₂—, —OC(O)—, —C(O)CO—,    —C(O)—, —C(O)NH—, —C(O)NR⁶—, —C(═N—CN), —NHCO—, —NR^(L)CO—,    —NHC(O)O—, —NR^(L)C(O)O—, —SO₂NH—, —SO₂NR^(L)—, —NHSO₂—,    —NR^(L)SO₂—, —NHC(O)NH—, —NR^(L)C(O)NH—, —NHC(O)NR^(L)—,    —NR^(L)C(O)NR^(L), —OC(O)NH—, —OC(O)NR^(L)—, —NHSO₂NH—,    —NR^(L)SO₂NH—, —NHSO₂NR^(L)—, —NR^(L)SO₂NR^(L), —SO— or —SO₂—;-   R^(L) is selected from C₁₋₆ aliphatic, C₃₋₁₀ cycloaliphatic, C₆₋₁₀    aryl, 5-10 membered heteroaryl or 5-10 membered heterocyclyl; or two    R^(L) groups, on the same substituent or different substituents,    together with the atom(s) to which each R^(L) group is bound, form a    3-8 membered heterocyclyl;-   each p is independently 0 or 1;-   Q² and Q³ are each independently selected from a bond or a C₁₋₆    alkylidene chain, wherein up to two methylene units of the chain are    each optionally and independently replaced by —NR′—, —S—, —O—, —CS—,    —CO₂—, —OCO—, —CO—, —COCO—, —CONR′—, —NR′CO—, —NR′CO₂—, —SO₂NR′—,    —NR′SO₂—, —CONR′NR′—, —NR′CONR′—, —OCONR′—, —NR′NR′—, —NR′SO₂NR′—,    —SO—, —SO₂—, —PO—, —PO₂—, or —POR′—; and wherein any carbon atom in    the one or more methylene units is optionally substituted with one    or two occurrences of R⁶, wherein each occurrence of R⁶ is    independently halogen, —CN, —NO₂, or —U_(n)R′, or two occurrences of    R⁶, or R′ and R⁶, taken together with the atoms to which they are    bound, form an optionally substituted 3-6-membered cycloalkyl,    heterocyclyl, aryl or heteroaryl ring; and-   Ar¹ and Ar² are each independently selected from a C₁₋₆ aliphatic, a    3-8 membered saturated, partially unsaturated, or fully unsaturated    monocyclic ring having 0-3 heteroatoms independently selected from    nitrogen, oxygen, or sulfur, or an 8-12 membered saturated,    partially unsaturated, or fully unsaturated bicyclic ring system    having 0-5 heteroatoms independently selected from nitrogen, oxygen,    or sulfur; wherein Ar¹ and Ar² are each optionally substituted with    0-5 independent occurrences of TR⁷; wherein T is a bond or is a    C₁-C₆ alkylidene chain wherein up to two methylene units of T are    optionally and independently replaced by —NR—, —S—, —O—, —CS—,    —CO₂—, —OCO—, —CO—, —COCO—, —CONR—, —NRCO—, —NRCO₂—, —SO₂NR—,    —NRSO₂—, —CONRNR—, —NRCONR—, —OCONR—, —NRNR—, —NRSO₂NR—, —SO—,    —SO₂—, —PO—, —PO₂—, or —POR—; and each occurrence of R⁷ is    independently selected from —R′, halogen, —NO₂, —CN or ═O.

In another embodiment, a compound of the invention has one of formulaeII-VII:

wherein R^(1Z1), R^(1Z2), R^(1Z3) and R^(1Z4) are each independentlyselected from H, halogen, —CN, —NO₂, or —V_(m)R′. In a furtherembodiment, the compound has one of formulae II, III or VI.

In one embodiment, the compound is any one formuale I-VII and

In a further embodiment, the compound is either of formulae II, III orVI. In another embodiment, X₁ is CR⁴ and X₂ is N or CR⁴. In a furtherembodiment, R⁴ is H.

In one embodiment, the compound is any one of formuale I-VII and R¹ isindependently selected from H, halogen or C₁₋₃ aliphatic. In a furtherembodiment, the compound is either of formulae II, III or VI and

In yet a further embodiment, R¹ is independently selected from H orhalogen.

In one embodiment of formulae II-VII, R^(1Z1), R^(1Z2), R^(1Z3) andR^(1Z4), if present, are each independently selected from H, halogen orC₁₋₃ aliphatic. In a further embodiment, R^(1Z1) and R^(1Z2), ifpresent, are H. In yet another embodiment, R^(1Z3) and R^(1Z4), ifpresent, are H or halogen. In a further embodiment, R^(1Z3) and R^(1Z4),if present, are H or F and at least one of R^(1Z3) and R^(1Z4) is H. Inyet a further embodiment, R^(1Z3) and R^(1Z4), if present, are H. Inanother embodiment, R^(1Z1), R^(1Z2), R^(1Z3) and R^(1Z4), if present,are H.

In one embodiment, the compound is any one of formulae I-VII and Q¹ is—CO—, —SO₂—, —NR², —NR²CO—, —CONR²—, —SO₂NR². In a further embodiment,the compound is either of formulae II, III or VI and

In a further embodiment, G is —NR² and Q¹ is —CO—, or G is —CO— and Q¹is —NR²—. In yet a further embodiment, R² is H, —C₁₋₄ aliphatic,-cyclopropyl, (CH₂)₁₋₃OH or

In a still further embodiment, R² is H.

In another embodiment of the invention, the compound is any one offormulae I-VII and R³ is Q²-Ar¹. In a further embodiment, the compoundis either of formulae II, III or VI and

In a further embodiment, Q² is —(CHR⁶)_(q)—, —(CHR⁶)_(q)O—,—(CHR⁶)_(q)S—, —(CHR⁶)_(q)S(O)₂—, —(CHR⁶)_(q)S(O)—, —(CHR⁶)_(q)NR—, or—(CHR⁶)_(q)C(O)—, wherein q is 0, 1, 2, or 3, and each R⁶ is R′,—N(R)(R′), —(CH₂)₁₋₄N(R)(R′), —(CH₂)₁₋₄C(CH₃)₂N(R)(R′),—(CH₂)₁₋₄CH(CH₃)N(R)(R′), —OR′, —(CH₂)₁₋₄OR′, —NR(CH₂)₁₋₄N(R)(R′),—NR(CH₂)₁₋₄SO₂R′, —NR(CH₂)₁₋₄COOR′, or —NR(CH₂)₁₋₄COR′, or twooccurrences of R⁶, taken together with the atoms to which they arebound, form an optionally substituted 3-6-membered saturated, partiallyunsaturated, or fully unsaturated ring. In yet a further embodiment, Q²is —(CHR⁶)_(q)—, q is 1 or 2, and R⁶ is R′, —N(R)(R′),—(CH₂)₁₋₄N(R)(R′), —OR′, —(CH₂)₁₋₄OR′ or —NR(CH₂)₁₋₄SO₂R′. In a stillfurther embodiment, Q² is —(CHR⁶)_(q)—, q is 1 or 2, and each R⁶ is H.

In another embodiment, Ar¹ is a C₃₋₆ aliphatic, a 5-8 memberedsaturated, partially unsaturated, or fully unsaturated monocyclic ringhaving 0-3 heteroatoms independently selected from nitrogen, oxygen, orsulfur, or an 8-12 membered saturated, partially unsaturated, or fullyunsaturated bicyclic ring system having 0-5 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur; wherein Ar¹ is optionallysubstituted with 0-5 independent occurrences of TR⁷.

In a further embodiment, Ar¹ is selected from one of

wherein t is 0, 1, 2, 3, 4, or 5, and wherein any Ar¹ is bonded to Qthrough any substitutable nitrogen or carbon atom, and wherein one ormore hydrogen atoms on any substitutable nitrogen or carbon atom issubstituted with one or more independent occurrences of TR⁷. In afurther embodiment, Ar¹ is

In yet a further embodiment, Ar¹ is

In some embodiments, t is 0, 1 or 2, and each TR⁷ is independentlyselected from halogen, —CN, —R′, —OR′, —NRR′, —OSO₂R′, —NRSO₂R′,—NRSO₂NRR′, —SO₂NRR′, —CONRR′, —COR′, —COOR′, —NRCOR′ or —SO₂R′. In afurther embodiment, TR⁷ is selected from —F, —Cl, —CN, —NH₂, —CH₃,—CH₂CH₃, —CH(CH₃)₂, —OR^(x), —OCF₃, —NR^(x)SO₂R^(x),—NR^(x)SO₂N(R^(x))₂, —COOC(CH₃)₃, —OSO₂CH₃, —OH, —SO₂N(R^(x))₂,—SO₂N(R^(x))₂, —SO₂R^(x), -pyrollidinone, tetrahydrofuran or-D-(CH₂)_(p)—Y, wherein R^(x) is a H or a C₁₋₄ alkyl, D is —SO₂—,—SO₂NH—, —NHSO₂— or —O—, p is 0-3, and Y is selected from:

wherein R^(y) is H or C₁₋₃ alkyl, and wherein one or more carbon atomsof Y is optionally substituted with ═O.

In one embodiment, t is 1 or 2 and one TR⁷ is -D-(CH₂)_(p)—Y, D is —O—,p is 2 or 3, Y is

and R^(y) is H or CH₃, and wherein one or more carbon atoms of Y isoptionally substituted with ═O. In another embodiment, t is 1 or 2 andone TR⁷ is —SO₂N(R^(x))₂, —NR^(x)SO₂R^(x), —NHSO₂R^(x), —OCF₃, or—OR^(x). In yet another embodiment, t is 1 or 2, and one or both TR⁷ areF or Cl. In yet another embodiment, Ar¹ is

and t is 0 or 1.

Representative examples of compounds of formula I are set forth below inTable 1 below. TABLE 1 Examples of Compounds of Formula I: 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

General Synthetic Methodology

The compounds of this invention may be prepared in general by methodsknown to those skilled in the art for analogous compounds, asillustrated by the general schemes below, and the preparative examplesthat follow.

Although certain exemplary embodiments are depicted and described aboveand herein, it will be appreciated that compounds of the invention canbe prepared according to the methods described generally above usingappropriate starting materials by methods generally available to one ofordinary skill in the art.

Uses, Formulation and Administration

As discussed above, the present invention provides compounds that areinhibitors of protein kinases, and thus the present compounds are usefulfor the treatment of diseases, disorders, and conditions including, butnot limited to a proliferative disorder, a cardiac disorder, aneurodegenerative disorder, psychotic disorders, an autoimmune disorder,a condition associated with organ transplant, an inflammatory disorder,an immunologically mediated disorder, a viral disease, or a bonedisorder. In preferred embodiments, the compounds are useful for thetreatment of hypertension, angina, angina pectoris, cerebrovascularcontraction, asthma, peripheral circulation disorder, premature birth,cancer, erectile dysfunction, arteriosclerosis, spasm (cerebralvasospasm and coronary vasospasm), retinopathy (e.g., glaucoma),inflammatory disorders, autoimmune disorders, AIDS, osteoporosis,myocardial hypertrophy, ischemia/reperfusion-induced injury, endothelialdysfunction, Alzheimer's disease, or benign prostatic hyperplasia. Inother embodiments, such conditions in which ROCK is known to play a roleinclude, without limitation, hypertension, cerebral vasospasm, coronaryvasospasm, bronchial asthma, preterm labor, erectile dysfunction,glaucoma, vascular smooth muscle cell proliferation, myocardialhypertrophy, malignoma, ischemia/reperfusion-induced injury, endothelialdysfunction, Crohn's Disease and colitis, neurite outgrowth, Raynaud'sDisease, angina, Alzheimer's disease, benign prostatic hyperplasia, oratherosclerosis.

Accordingly, in another aspect of the present invention,pharmaceutically acceptable compositions are provided, wherein thesecompositions comprise any of the compounds as described herein, andoptionally comprise a pharmaceutically acceptable carrier, adjuvant orvehicle. In certain embodiments, these compositions optionally furthercomprise one or more additional therapeutic agents.

It will also be appreciated that certain of the compounds of presentinvention can exist in free form for treatment, or where appropriate, asa pharmaceutically acceptable derivative thereof. According to thepresent invention, a pharmaceutically acceptable derivative includes,but is not limited to, pharmaceutically acceptable prodrugs, salts,esters, salts of such esters, or any other adduct or derivative whichupon administration to a patient in need is capable of providing,directly or indirectly, a compound as otherwise described herein, or ametabolite or residue thereof.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgement,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. A“pharmaceutically acceptable salt” means any non-toxic salt or salt ofan ester of a compound of this invention that, upon administration to arecipient, is capable of providing, either directly or indirectly, acompound of this invention or an inhibitorily active metabolite orresidue thereof. As used herein, the term “inhibitorily activemetabolite or residue thereof” means that a metabolite or residuethereof is also an inhibitor of a ROCK kinase.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge et al., describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporatedherein by reference. Pharmaceutically acceptable salts of the compoundsof this invention include those derived from suitable inorganic andorganic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. This inventionalso envisions the quaternization of any basic nitrogen-containinggroups of the compounds disclosed herein. Water or oil-soluble ordispersable products may be obtained by such quaternization.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

As described above, the pharmaceutically acceptable compositions of thepresent invention additionally comprise a pharmaceutically acceptablecarrier, adjuvant, or vehicle, which, as used herein, includes any andall solvents, diluents, or other liquid vehicle, dispersion orsuspension aids, surface active agents, isotonic agents, thickening oremulsifying agents, preservatives, solid binders, lubricants and thelike, as suited to the particular dosage form desired. Remington'sPharmaceutical Sciences, Sixteenth Edition, E. W. Martin (MackPublishing Co., Easton, Pa., 1980) discloses various carriers used informulating pharmaceutically acceptable compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, or potassiumsorbate, partial glyceride mixtures of saturated vegetable fatty acids,water, salts or electrolytes, such as protamine sulfate, disodiumhydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zincsalts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, woolfat, sugars such as lactose, glucose and sucrose; starches such as cornstarch and potato starch; cellulose and its derivatives such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; powderedtragacanth; malt; gelatin; talc; excipients such as cocoa butter andsuppository waxes; oils such as peanut oil, cottonseed oil; saffloweroil; sesame oil; olive oil; corn oil and soybean oil; glycols; such apropylene glycol or polyethylene glycol; esters such as ethyl oleate andethyl laurate; agar; buffering agents such as magnesium hydroxide andaluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline;Ringer's solution; ethyl alcohol, and phosphate buffer solutions, aswell as other non-toxic compatible lubricants such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releasingagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

In yet another aspect, a method for the treatment or lessening theseverity of a proliferative disorder, a cardiac disorder, aneurodegenerative disorder, a psychotic disorder, an autoimmunedisorder, a condition associated with organ transplant, an inflammatorydisorder, an immunologically mediated disorder, a viral disease, or abone disorder is provided comprising administering an effective amountof a compound, or a pharmaceutically acceptable composition comprising acompound to a subject in need thereof. In certain embodiments of thepresent invention an “effective amount” of the compound orpharmaceutically acceptable composition is that amount effective fortreating or lessening the severity of a proliferative disorder, acardiac disorder, a neurodegenerative disorder, a psychotic disorder, anautoimmune disorder, a condition associated with organ transplant, aninflammatory disorder, an immunologically mediated disorder, a viraldisease, or a bone disorder. The compounds and compositions, accordingto the method of the present invention, may be administered using anyamount and any route of administration effective for treating orlessening the severity of a proliferative disorder, a cardiac disorder,a neurodegenerative disorder, an autoimmune disorder, a conditionassociated with organ transplant, an inflammatory disorder, animmunologically mediated disorder, a viral disease, or a bone disorder.The exact amount required will vary from subject to subject, dependingon the species, age, and general condition of the subject, the severityof the infection, the particular agent, its mode of administration, andthe like. The compounds of the invention are preferably formulated indosage unit form for ease of administration and uniformity of dosage.The expression “dosage unit form” as used herein refers to a physicallydiscrete unit of agent appropriate for the patient to be treated. Itwill be understood, however, that the total daily usage of the compoundsand compositions of the present invention will be decided by theattending physician within the scope of sound medical judgment. Thespecific effective dose level for any particular patient or organismwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The term “patient”, as usedherein, means an animal, preferably a mammal, and most preferably ahuman.

The pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated. Incertain embodiments, the compounds of the invention may be administeredorally or parenterally at dosage levels of about 0.01 mg/kg to about 50mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subjectbody weight per day, one or more times a day, to obtain the desiredtherapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar—agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, and eye drops are also contemplatedas being within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

As described generally above, the compounds of the invention are usefulas inhibitors of protein kinases. In one embodiment, the compounds andcompositions of the invention are inhibitors of ROCK, and thus, withoutwishing to be bound by any particular theory, the compounds andcompositions are particularly useful for treating or lessening theseverity of a disease, condition, or disorder where activation of ROCKis implicated in the disease, condition, or disorder. When activation ofROCK is implicated in a particular disease, condition, or disorder, thedisease, condition, or disorder may also be referred to as“ROCK-mediated disease” or disease symptom. Accordingly, in anotheraspect, the present invention provides a method for treating orlessening the severity of a disease, condition, or disorder whereactivation of ROCK is implicated in the disease state.

The activity of a compound utilized in this invention as an inhibitor ofROCK, may be assayed in vitro, in vivo or in a cell line. In vitroassays include assays that determine inhibition of either thephosphorylation activity or ATPase activity of activated ROCK. Alternatein vitro assays quantitate the ability of the inhibitor to bind to ROCK.Inhibitor binding may be measured by radiolabelling the inhibitor priorto binding, isolating the inhibitor/ROCK complex and determining theamount of radiolabel bound. Alternatively, inhibitor binding may bedetermined by running a competition experiment where new inhibitors areincubated with ROCK bound to known radioligands.

The term “measurably inhibit”, as used herein means a measurable changein ROCK activity between a sample comprising said composition and ROCKand an equivalent sample comprising ROCK in the absence of saidcomposition.

The term “ROCK-mediated condition” or “disease”, as used herein, meansany disease or other deleterious condition in which ROCK is known toplay a role. The term “ROCK-mediated condition” or “disease” also meansthose diseases or conditions that are alleviated by treatment with aROCK inhibitor. Such conditions include, without limitation,hypertension, angina, angina pectoris, cerebrovascular contraction,asthma, peripheral circulation disorder, premature birth, cancer,erectile dysfunction, arteriosclerosis, spasm (cerebral vasospasm andcoronary vasospasm), retinopathy (e.g., glaucoma), inflammatorydisorders, autoimmune disorders, AIDS, osteoporosis, myocardialhypertrophy, ischemia/reperfusion-induced injury, endothelialdysfunction, Alzheimer's disease, or benign prostatic hyperplasia. Inother embodiments, such conditions in which ROCK is known to play a roleinclude, without limitation, hypertension, cerebral vasospasm, coronaryvasospasm, bronchial asthma, preterm labor, erectile dysfunction,glaucoma, vascular smooth muscle cell proliferation, myocardialhypertrophy, malignoma, ischemia/reperfusion-induced injury, endothelialdysfunction, Crohn's Disease and colitis, neurite outgrowth, Raynaud'sDisease, angina, Alzheimer's disease, benign prostatic hyperplasia, oratherosclerosis.

It will also be appreciated that the compounds and pharmaceuticallyacceptable compositions of the present invention can be employed incombination therapies, that is, the compounds and pharmaceuticallyacceptable compositions can be administered concurrently with, prior to,or subsequent to, one or more other desired therapeutics or medicalprocedures. The particular combination of therapies (therapeutics orprocedures) to employ in a combination regimen will take into accountcompatibility of the desired therapeutics and/or procedures and thedesired therapeutic effect to be achieved. It will also be appreciatedthat the therapies employed may achieve a desired effect for the samedisorder (for example, an inventive compound may be administeredconcurrently with another agent used to treat the same disorder), orthey may achieve different effects (e.g., control of any adverseeffects). As used herein, additional therapeutic agents that arenormally administered to treat or prevent a particular disease, orcondition, are known as “appropriate for the disease, or condition,being treated”.

For example, chemotherapeutic agents or other anti-proliferative agentsmay be combined with the compounds of this invention to treatproliferative diseases and cancer. Examples of known chemotherapeuticagents include, but are not limited to, For example, other therapies oranticancer agents that may be used in combination with the inventiveanticancer agents of the present invention include surgery, radiotherapy(in but a few examples, gamma.-radiation, neutron beam radiotherapy,electron beam radiotherapy, proton therapy, brachytherapy, and systemicradioactive isotopes, to name a few), endocrine therapy, biologicresponse modifiers (interferons, interleukins, and tumor necrosis factor(TNF) to name a few), hyperthermia and cryotherapy, agents to attenuateany adverse effects (e.g., antiemetics), and other approvedchemotherapeutic drugs, including, but not limited to, alkylating drugs(mechlorethamine, chlorambucil, Cyclophosphamide, Melphalan,Ifosfamide), antimetabolites (Methotrexate), purine antagonists andpyrimidine antagonists (6-Mercaptopurine, 5-Fluorouracil, Cytarabile,Gemcitabine), spindle poisons (Vinblastine, Vincristine, Vinorelbine,Paclitaxel), podophyllotoxins (Etoposide, Irinotecan, Topotecan),antibiotics (Doxorubicin, Bleomycin, Mitomycin), nitrosoureas(Carmustine, Lomustine), inorganic ions (Cisplatin, Carboplatin),enzymes (Asparaginase), and hormones (Tamoxifen, Leuprolide, Flutamide,and Megestrol), Gleevec™, adriamycin, dexamethasone, andcyclophosphamide. For a more comprehensive discussion of updated cancertherapies see, http://www.nci.nih.gov/, a list of the FDA approvedoncology drugs at http://www.fda.gov/cder/cancer/druglistframe.htm, andThe Merck Manual, Seventeenth Ed. 1999, the entire contents of which arehereby incorporated by reference.

Other examples of agents the inhibitors of this invention may also becombined with include, without limitation: treatments for Alzheimer'sDisease such as Aricept® and Excelon®; treatments for Parkinson'sDisease such as L-DOPA/carbidopa, entacapone, ropinrole, pramipexole,bromocriptine, pergolide, trihexephendyl, and amantadine; agents fortreating Multiple Sclerosis (MS) such as beta interferon (e.g., Avonex®and Rebif®), Copaxone®, and mitoxantrone; treatments for asthma such asalbuterol and Singulair®; agents for treating schizophrenia such aszyprexa, risperdal, seroquel, and haloperidol; anti-inflammatory agentssuch as corticosteroids, TNF blockers, IL-1 RA, azathioprine,cyclophosphamide, and sulfasalazine; immunomodulatory andimmunosuppressive agents such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, interferons, corticosteroids, cyclophosphamide,azathioprine, and sulfasalazine; neurotrophic factors such asacetylcholinesterase inhibitors, MAO inhibitors, interferons,anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonianagents; agents for treating cardiovascular disease such asbeta-blockers, ACE inhibitors, diuretics, nitrates, calcium channelblockers, and statins; agents for treating liver disease such ascorticosteroids, cholestyramine, interferons, and anti-viral agents;agents for treating blood disorders such as corticosteroids,anti-leukemic agents, and growth factors; and agents for treatingimmunodeficiency disorders such as gamma globulin.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered in a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

The compounds of this invention or pharmaceutically acceptablecompositions thereof may also be incorporated into compositions forcoating implantable medical devices, such as prostheses, artificialvalves, vascular grafts, stents and catheters. Accordingly, the presentinvention, includes a composition for coating an implantable devicecomprising a compound of the present invention as described generallyabove, and in classes and subclasses herein, and a carrier suitable forcoating said implantable device. In still another aspect, the presentinvention includes an implantable device coated with a compositioncomprising a compound of the present invention as described generallyabove and a carrier suitable for coating said implantable device.

Vascular stents, for example, have been used to overcome restenosis.However, patients using stents or other implantable devices risk clotformation or platelet activation. These unwanted effects may beprevented or mitigated by pre-coating the device with a pharmaceuticallyacceptable composition comprising a kinase inhibitor. Suitable coatingsand the general preparation of coated implantable devices are describedin U.S. Pat. Nos. 6,099,562; 5,886,026; and 5,304,121. The coatings aretypically biocompatible polymeric materials such as a hydrogel polymer,polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylacticacid, ethylene vinyl acetate, and mixtures thereof. The coatings mayoptionally be further covered by a suitable topcoat of fluorosilicone,polysaccarides, polyethylene glycol, phospholipids or combinationsthereof to impart controlled release characteristics in the composition.

Another aspect of the invention relates to inhibiting ROCK activity in abiological sample or a patient, which method comprises administering tothe patient, or contacting said biological sample with a compound offormula I or a composition comprising said compound. The term“biological sample”, as used herein, includes, without limitation, cellcultures or extracts thereof; biopsied material obtained from a mammalor extracts thereof; and blood, saliva, urine, feces, semen, tears, orother body fluids or extracts thereof. Inhibition of ROCK kinaseactivity in a biological sample is useful for a variety of purposes thatare known to one of skill in the art. Examples of such purposes include,but are not limited to, blood transfusion, organ-transplantation,biological specimen storage, and biological assays.

All references provided in the Examples are herein incorporated byreference. As used herein, all abbreviations, symbols and conventionsare consistent with those used in the contemporary scientificliterature. See, e.g., Janet S. Dodd, ed., The ACS Style Guide: A Manualfor Authors and Editors, 2nd Ed., Washington, D.C.: American ChemicalSociety, 1997, herein incorporated in its entirety by reference.

EXAMPLES Example 1 Preparation of Compound 9

4-Bromothiophene-2-carboxylic acid (A)

A solution of sodium chlorite (47.5 g, 525 mmol) and sodium hydrogenphosphate (63.0 g, 525 mmol) in water (500 mL) was added slowly to astirred solution of 4-bromo-2-thiophenecarboxaldehyde (50 g, 261 mmol)in tertiary butanol (600 mL) and 2-methylbutene (8 mL) in 30 min at 0°C. The cooling bath was removed and the resulting solution was stirredat room temperature for 16 h. The mixture was transferred to a separatorfunnel and the aqueous layer was separated. Organic layer (tertiarybutanol) was concentrated under reduced pressure to give white residue,which was added to the aqueous aliquot. The total aqueous layer wasacidified with 6N HCl (100 mL). The precipitated product was extractedwith ethyl acetate (3×250 mL). The organic layer was dried (Na₂SO₄) andconcentrated under reduced pressure to afford the title4-bromothiophene-2-carboxylic acid (A) as a white solid (54 g, 100%).FIA MS 207 (M+1).

tert-Butyl-4-bromothiophen-2-yl-carbamate (B)

A mixture of 4-bromothiophene-2-carboxylic acid (A) (53 g, 255 mmol),diphenylphosphoryl azide (Aldrich, 70 mL, 323 mmol), triethylamine (45mmol) in tertiary butanol (675 mL) was heated at 100° C. for 5 h andthen cooled to room temperature. The solvent was evaporated to givebrown gum, which was dissolved in EtOAc (500 mL). The organic solutionwas washed with saturated NaHCO₃ (500 mL) and water (500 mL)respectively, then dried over Na₂SO₄ and concentrated. The crude productwas dissolved in CH₂Cl₂ (75 mL) and purified by flash columnchromatography on silica gel (10%-15% EtOAc/hexanes) to afford titlecompound tert-butyl-4-bromothiophen-2-yl-carbamate (B) (47 g, 66%) as awhite solid. FIA MS 278 (M+1).

5-BOC^(t)aminothiophen-3-yl-3-boronic acid (C)

2.5M nBuLi in hexane (7.2 mL, 18 mmol) was added dropwise to a stirredsolution of tert-butyl-4-bromothiophen-2-yl-carbamate (B) (1 g, 3.6mmol) in THF (3 mL) and toluene (11 mL) at −78° C. under nitrogen andthe solution was stirred 1 h at −78° C.

Triisopropylborate (2 mL, 9 mmol) was added and the resulting thickbrown solution was stirred at −78° C. for 30 min and room temperaturefor 30 min. 2N HCl (15 ml) and ethyl acetate (50 mL) were added and thesolution was stirred 25 min. The organic layer was separated and aqueouslayer was extracted with EtOAc (2×25 mL). The combined organic layerswere dried (Na₂SO₄) and concentrated to give title compound5-aminothiophen-3-yl-3-boronic acid (C) as an oil (0.8 g, 91%). Theproduct was used for next step without purification. FIA MS 242 (M−1).

5-Aminothiophen-3-yl-3-boronic acid (D)

The crude 5-aminothiophen-3-yl-3-boronic acid (C) (0.8 g) was dissolvedin 4N HCl (10 mL) and methanol (2 mL) and the solution was stirred atroom temperature for 2 h. The solvent was evaporated under reducedpressure to give title compound 5-BOC^(t)aminothiophen-3-yl-3-boronicacid (D) (0.58 g) as a brown gum. The product was used for next stepwithout purification. FIA MS 143 (M+1).

5-(2-(3-Methsulfonamidophenyl)acetamido)thiophen-3-yl-3-boronic acid (E)

A mixture of 5-aminothiophen-3-yl-3-boronic acid (D) (0.48 g, 3.35mmol), 3-methylsulfonamidephenylacetic acid (0.77 g, 3.35 mmol),BtSO₂CH₃ (0.66 g, 3.35 mmol), Et₃N (3 mL) in THF (25 mL) and DMF (2 mL)was heated at 85° C. for 16 h and then cooled to room temperature. Thesolvent was evaporated and the crude material was dissolved in EtOAc (50mL). The organic solution was washed with saturated NaHCO₃ (20 mL),brine (20 mL), dried (Na₂SO₄) and concentrated. The crude material waspurified by chromatography. The impurities were first eluted with 40-70%EtOAc/hexanes and the product was eluted with 5-30% MeOH/CH₂Cl₂. Yield0.54 g, 46%, FIA MS 355 (M+1).

5-(2-(5-Methsulfonamido-2-fluorophenyl)acetamido)thiophen-3-yl-3-boronicacid (F)

The boronic acid F was synthesized and purified in a manner similar to Eusing a mixture of D (0.48 g, 3.35 mmol),3-methylsulfonamide-6-flurophenylacetic acid (0.82 g, 3.35 mmol),BtSO₂CH₃ (0.66 g, 3.35 mmol), Et₃N (3 mL). Yield 0.43 g, 34%, FIA MS 373(M+1).

5-(2-(3-N,N-dimethylaminosulfonylbenzene)acetamido)thiophen-3-yl-3-boronicacid (G)

The boronic acid F was synthesized and purified in a manner similar to Eusing a mixture of D (0.48 g, 3.35 mmol), 3-N,N-dimethylsulfonephenylacetic acid (0.81 g, 3.35 mmol), BtSO₂CH₃ (0.66 g, 3.35 mmol),Et₃N (3 mL). Yield 0.64 g, 52%, FIA MS 369 (M+1).

5-(2-(3-(3-(tert-butoxycarbonyl-piperidin-4-yl)propoxy)phenyl)acetamido)thiophene-3-yl-3-boronicacid (H)

The boronic acid H was synthesized and purified in a manner similar to Eusing a mixture of D (0.48 g, 3.35 mmol),2-(3-(3-(BOC^(t)piperidin-4-yl)propoxy)phenylacetic acid (1.26 g, 3.35mmol), BtSO₂CH₃ (0.66 g, 3.35 mmol), Et₃N (3 mL). Yield 0.57 g, 43.6%,FIA MS 503 (M+1).

N-(4-(1H-pyrazolo[3,4-b]pyrazin-3-yl)thiophen-2-yl)-2-(5-methsulfonamido-2-fluorophenyl)acetamide(Compound 9)

A mixture of J (0.050 g, 0.134 mmol), M (see Monatshefte fur Chemie113:731, 1992; 0.026 g, 0.134 mmol), Pd₂(dba)₃ (0.017 g, 0.02 mmol),P(Bu^(t))₃ (0.01 g, 0.04 mmol), KF.H₂O (0.1 g, 1.3 mmol) in 1,4-dioxane(2 mL) and water (0.5 mL) was heated at 185° C. in a microwave for 25min. Solid was filtered and washed with EtOAc (1 mL). The filtrate wasconcentrated and the crude product was purified by preparative HPLC(5-75% CH₃CN/water, 15 min) to give title compound 9 (0.005 g) as anoil.

Example 2 Preparation of Compounds 6, 7, 8 and 10

1 Fluropyridine-3-carbaldehyde (N)

Under nitrogen, 2.5M nBuLi in hexane (25.6 mL, 64.37 mmol) was addeddropwise to a stirred solution of isopropylamine (9 mL, 64 mmol) in THF(50 mL) and at −78° C. for 10 min. and the solution was stirred 30 minat −78° C. 2-Fluropyridine (4.4 mL, 51.2 mmol) was added and thesolution was stirred at −78° C. for 2 h. A cold solution ofN-methyl-N-(pyridin-2-yl)formamide (7 g, 51.2 mmol) in THF (30 mL) wereadded in 15 min. The resulting solution was warmed to room temperatureand stirred for 1 h. 2N HCl (15 mL) and ethyl acetate (50 mL) ware addedand the solution was stirred 25 min. The organic layer was separated andaqueous layer was extracted with EtOAc (2×25 mL). The combined organiclayers were dried (Na₂SO₄) and concentrated. The crude product waspurified by chromatography (10-60% EtOAc/hexanes) to give title compoundN (1.3 g, 20%) as a yellow oil. ¹H NMR (DMSO-d6, 500 MHz) δ 10.16(s,1H), 8.53(dd, 1H), 8.38(m, 1H), 7.58(m, 1H).

(E)-N′-((2-Fluoropyridin-3-yl)methylene)acetohydrazide (O)

A solution of N (1.3 g, 10.4 mmol), acylhydrazine (1.5 g, 20.8 mmol) inethanol (10 mL) was stirred at room temperature for 18 h. Theprecipitate was filtered and washed with ethanol (2×10 mL) and dried togive title compound O (0.82 g, 43%) as a white solid FIA MS 180 (M−1).

1H-Pyrazolo[3,4-b]pyridine (P)

A suspension of O (0.82 g, 4.5 mmol) in hydrazine monohydrate (5 mL) washeated at 90° C. for 15 min and poured into ice/water and extracted withethyl acetate (3×25 mL). The organic layers were dried (Na₂SO₄) andconcentrated to give title compound P (0.41 g, 77%) as a white solid. ¹HNMR (DMSO-d₆, 500 MHz) δ 13.60(brs, 1H), 8.55(d, 1H), 8.51(dd, 1H),8.14(s, 1H), 7.18(dd, 1H); FIA MS 120 (M+1).

3-Bromo-1H-pyrazolo[3,4-b]pyridine (Q)

Bromine (0.3 mL, 5.16 mmol) was added via a micro syringe to a stirredsolution of P (0.41 g, 3.45 mmol) in chloroform (10 mL). The resultingsuspension was stirred at room temperature for 2 h. The solvent wasevaporated and ethyl acetate (50 mL) was added. The organic layer waswashed with saturated K₂CO₃ and the organic layer was concentratedwithout drying at 70° C. to produce title compound Q (0.63 g, 92%). ¹HNMR (DMSO-d₆, 500 MHz) δ 8.56(t, 1H), 8.05(d, 1H), 7.24(m, 1H); FIA MS198(M−1).

3-Bromo-1-tosyl-1H-pyrazolo[3,4-b]pyridine (R)

Sodium hydride (0.05 g, 2 mmol) was added to a stirred solution of Q(0.1 g, 0.5 mmol) in THF (5 mL). The solution was stirred at roomtemperature for 30 min and P-TsCl (0.1 g, 0.5 mmol) was added. Theresulting suspension was heated at 60° C. for 45 min. and poured intowater (25 mL). The solution was extracted with ethyl acetate (2×25 mL),dried and concentrated to give title compound R (0.14, 80%). The productwas unstable and therefore used for next step immediately. FIA-MS353(M−1).

2-(2-Fluoro-5-methanesulfonylamino-phenyl)-N-[4-(1H-pyrazolo[3,4-b]pyridin-3-yl)-thiophen-2-yl]-acetamide(Compound 6).

A mixture of R (0.050 g, 0.142 mmol), F (0.053 g, 0.142 mmol), Pd₂(dba)₃(0.017 g, 0.02 mmol), P(Bu^(t))₃ (0.01 g, 0.04 mmol), KF.H₂O (0.1 g, 1.3mmol) in 1,4-dioxane (2 mL) and water (0.5 mL) was heated at 185° C. ina microwave for 25 min. Solid was filtered and washed with EtOAC (1 mL).The filtrate was concentrated and the crude product was purified bypreparative HPLC (5-75% CH₃CN/water, 15 min) to give title compound 6(0.027 g) as an oil.

2-(3-Methanesulfonylamino-phenyl)-N-[4-(1H-pyrazolo[3,4-b]pyridin-3-yl)-thiophen-2-yl]-acetamide(Compound 8).

The title compound 8 was synthesized and purified in a manner similar to6 using a mixture of R (0.050 g, 0.142 mmol), E (0.050 g, 0.142 mmol),Pd₂(dba)₃ (0.017 g, 0.02 mmol), P(Bu^(t))₃ (0.01 g, 0.04 mmol), KF.H₂O(0.1 g, 1.3 mmol) in 1,4-dioxane (2 mL) and water (0.5 mL). Yield 0.03g.

2-(3-Dimethylsulfamoyl-phenyl)-N-[4-(1H-pyrazolo[3,4-b]pyridin-3-yl)-thiophen-2-yl]-acetamide(Compound 10)

The title compound 10 was synthesized and purified in a manner similarto 6 using a mixture of R (0.050 g, 0.142 mmol), G (0.052 g, 0.142mmol), Pd₂(dba)₃ (0.017 g, 0.02 mmol), P(Bu^(t))₃ (0.01 g, 0.04 mmol),KF.H₂O (0.1 g, 1.3 mmol) in 1,4-dioxane (2 mL) and water (0.5 mL). Yield0.026 g.

2-(3-(3-Piperidin-4-yl-propoxy)-phenyl)-N-[4-(1H-pyrazolo[3,4-b]pyridin-3-yl)-thiophen-2-yl]-acetamide(Compound 7)

A mixture of R (0.050 g, 0.142 mmol), H (0.064 g, 0.142 mmol), Pd₂(dba)₃(0.017 g, 0.02 mmol), P(Bu^(t))₃ (0.01 g, 0.04 mmol), KF.H₂O (0.1 g, 1.3mmol) in 1,4-dioxane (2 mL) and water (0.5 mL) was heated at 185° C. ina microwave for 25 min. Solid was filtered and washed with EtOAC (1 mL).The filtrate was concentrated. The crude product was dissolved in CH₂Cl₂(1 mL) and TFA (1 mL). The solution was stirred at room temperature for1 h. and concentrated. The crude product was purified by preparativeHPLC (5-75% CH₃CN/water, 15 min) to give title compound 7 (0.032 g) asan oil.

Example 3 Preparation of Compounds 19 and 20

5-Bromo-3-trimethylsilanylethynyl-pyrazin-2-ylamine (U)

Trimethylsilylacetylene (3.4 mL) was added to a stirred suspension of T(see J. Heterocyclic. Chem. 1982, 19,673; 6.3 g, 25.12 mmol),Pd(PPh₃)₂Cl₂ (1.7 g), CuI (0.95 g) in triethyamine (20 mL) and THF (50mL) at room temperature and the solution was heated at 45° C. for 2 h.The solid was filtered off and the filtration was concentrated to givedark brown liquid. The crude product was purified by Biotage Horizon™eluting with 10%-50% EtOAc/hexane to afford title compound U (4.1 g,60%) as a yellow solid. ¹H NMR (DMSO-d₆, 500 MHz) δ 8.11(s, 1H),6.77(brs, 2H), 7.24(m, 1H); FIA MS 270(M+1).

2-Bromo-5H-pyrrolo[2,3-b]pyrazine (V)

A solution of U (4.0 g, 14.81 mmol) in THF (25 mL) was added to astirred suspension of potassium tert butoxide (2.5 g, 22.2 mmol) in THF(50 mL). The solution was refluxed for 3 h and then stirred at roomtemperature for 16 h. The solvent was evaporated and the residue wasdissolved in water (100 mL) and aqueous layer was extracted with ethylacetate (5×50 mL). Organic layer was dried and concentrated to givetitle compound V (1.62 g) as a yellow solid. ¹H NMR (DMSO-d6, 500 MHz) δ12.33(s, 1H), 8.35(s, 1H), 7.96(t, 1H), 6.63(dd, 1H); FIA MS 198(M−1).

5H-Pyrrolo[2,3-b]pyrazine (W)

10% Palladium on carbon (1.4 g) was added to a stirred, nitrogen flushedsolution of V (1.4 g, 7.14 mmol) and ammonium formate (4.2 g) in ethanol(50 mL). The solution was refluxed for 1 h and cooled to roomtemperature. The filtrate was concentrated to give a solid, which waswashed with ethyl acetate (3×20 mL). The solvent was evaporated and thecrude product was purified by Biotage Horizon eluting with 40%-80%EtOAC/hexane to afford title compound W (0.313 g, 37%) as a white solid.¹H NMR (DMSO-d6, 500 MHz) δ 12.05(s, 1H), 8.37(d, 1H), 8.22(d, 1H),7.85(d, 1H), 6.62(d, 1H); FIA MS 120(M+1).

7-Iodo-5H-pyrrolo[2,3-b]pyrazine (X)

1M Solution iodine monochloride (5.26 mL, 5.26 mmol) was added to astirred solution of W (0.313 g, 2.63 mmol) in CH₂Cl₂ (25 mL) and thesolution was stirred at room temperature for 16 h. The precipitatedyellow solid was filtered and suspended in ethyl acetate (50 mL) andwashed with saturated NaHCO₃ (25 mL). The organic layer was dried andconcentrated to give title compound 22 (0.64 g, 100%). ¹H NMR (DMSO-d6,500 MHz) δ 12.48(s, 1H), 8.45(d, 1H), 8.28(d, 1H), 8.10(d, 1H), 7.85(d,1H), FIA MS 246(M+1).

7-Iodo-5-(toluene-4-sulfonyl)-5H-pyrrolo[2,3-b]pyrazine (Y)

Sodium hydride (0.04 g, 1.68 mmol) was added to a stirred solution of X(0.103 g, 0.42 mmol) in THF (10 mL). The solution was stirred at roomtemperature for 30 min and P-TsCl (0.81 g, 0.62 mmol) was added. Theresulting suspension was heated a 60° C. for 45 min. and poured intowater (10 mL). The solution was extracted with ethyl acetate (2×25 mL),dried and concentrated to give title compound Y (0.168, 85%). FIA MS400(M+1).

2-(3-(3-Piperidin-4-yl-propoxy)-phenyl)-N-[4-(5H-pyrralo[2,3-b]pyrazin-3-yl)-thiophen-2-yl]-acetamide(Compound 19).

A mixture of Y (0.044 g, 0.11 mmol), H (0.055 g, 0.11 mmol), Pd₂(dba)₃(0.017 g, 0.02 mmol), P(Bu^(t))₃ (0.01 g, 0.04 mmol), KF.H₂O (0.1 g, 1.3mmol) in 1,4-dioxane (2 mL) and water (0.5 mL) was heated at 185° C. ina microwave oven for 25 min. Solid was filtered and washed with EtOAc (1mL). The filtrate was concentrated. The crude product was dissolved inCH₂Cl₂ (1 mL) and TFA (1 mL). The solution was stirred at roomtemperature for 1 h and concentrated. The crude product was purified bypreparative HPLC (5-75% CH₃CN/water, 15 min) to give title compound 19(0.0062 g) as an oil.

2-(3-Methanesulfonylamino-phenyl)-N-[5H-pyrralo[2,3-b]pyrazin-3-yl)-thiophen-2-yl]-acetamide(Compound 20).

The title compound 20 was synthesized and purified in a manner similarto 6 using a mixture of Y (0.050 g, 0.125 mmol), E (0.044 g, 0.125mmol), Pd₂(dba)₃ (0.017 g, 0.02 mmol), P(Bu^(t))₃ (0.01 g, 0.04 mmol),KF.H₂O (0.1 g, 1.3 mmol) in 1,4-dioxane (2 mL) and water (0.5 mL).

Example 4 Preparation ofN-(4-(1H-pyrazolo[3,4-b]pyridin-3-yl)thiazol-2-yl)-2-phenylacetamides

t-Butyl 2-(2-fluoropyridin-3-yl)-2-oxoacetate (BB)

To a solution of 2-fluoropyridine (5.82 g, 0.0599 mol) in 150 mL THF at−78° C., LDA (33.0 ml of 2M, 0.066 mol) was added dropwise and thereaction mixture was stirred at −78° C. for 2 h. This solution wastransferred slowly into chilled to −78° C. THF (500 ml) solution oft-butyl α-oxo-1 H-imidazole-1-acetate (AA) (see J. Org. Chem. 46:211,1981; 14.69 g, 0.0749 mol) by means of cannula under nitrogen pressure.The combined solution was continuously stirred at −78° C. for 30 min.The reaction mixture was poured into sat. NH₄Cl (500 mL). The organicphase was separated and dried (MgSO₄). After removing solvent, theproduct was purified by chromatography to give title compound BB (6.7 g,49.6% yield). H NMR (CDCl₃): 1.6(s, 9H), 7.4(dd, 1H), 8.36(dd, 1H),8.5(dd, 1H).

tert-Butyl 2-(2-fluoropyridin-3-yl)-2-hydrazonoacetate (CC)

To a solution of t-butyl 2-(2-fluoropyridin-3-yl)-2-oxoacetate (BB) inmethylene dichloride at room temperature, titanium(IV) isopropoxide wasadded, followed by hydrazine hydrate dropwise. The reaction mixture wasstirred at room temperature for 10 h, then 6 mL water was added. Thereaction mixture was continuously stirred at room temperature forovernight. The reaction mixture was filtered and the filtration cake waswashed with methylene chloride 3 times. After removal of solvent, thefiltrate was purified by chromatography 95%/5% methylenedichloride/methanol to give the title compound CC (5.0 g, 70% yield). HNMR (CDCl₃): 1.5(s, 9H), 6.18(s, br, 2H), 7.3(dd, 1H), 7.74(dd, 1H),8.3(dd, 1H).

tert-Butyl 1H-pyrazolo[3,4]pyridine-3-carboxylate (DD)

To a solution of t-butyl 2-(2-fluoropyridin-3-yl)-2-hydrazonoacetate(CC) (0.58 g, 2.42 mmol) in THF, sodium hydride (0.116 g, 60% mineraloil dispersion, 2.9 mmol) was added in portions. The reaction mixturewas stirred at room temperature until no further gas was released, thenwarmed to 50° C. for 2 h. Ethyl acetate and brine was added to thereaction mixture and the organic layer was separated and dried overMgSO₄. After removal of the solvent, the product was purified bychromatography with 50%/50% ethyl acetate/hexanes to give title compoundDD (0.4 g, 75% yield). H NMR (CDCl₃): 1.67(s, 9H), 7.28(dd, 1H),8.52(dd, 1H), 8.63(dd, 1H).

1H-Pyrazolo[3,4-b]pyridine-3-carboxylic acid (EE)

A THF/CH₂Cl₂ solution of tert-butyl1H-pyrazolo[3,4]pyridine-3-carboxylate (DD) (3.0 g, 13.7mmol) wasstirred at room temperature for 3 h. The solvent was removed to givetitle compound EE (2.2 g, 99% yield). H NMR (DMSO): 7.38(dd, 1H),8.45(dd, 1H), 8.63(dd, 1H).

1H-Pyrazolo[3,4-b]pyridine-3-carbonyl chloride (FF)

Thionyl chloride, chloroform and a catalytic amount of DMF was added toa solution of 1H-pyrazolo[3,4-b]pyridine-3-carboxylic acid (EE) (5.6 g,34.3 mmol) and the mixture was refluxed overnight. The reaction mixturewas cooled to room temperature and filtered. The filtration cake waswashed with ether to give the title compound FF.

2-(1H-Pyrazolo[3,4-b]pyridine-3-carbonyl)-malonic acid dimethyl ester(GG)

Sodium hydride was added in portions to a dimethylmelanate (0.33 g, 2.5mmol)/THF solution at 0° C., then the reaction mixture was refluxed for1 h. A THF suspension of 1H-Pyrazolo[3,4-b]pyridine-3-carbonyl chloride(FF) was added. The reaction mixture was refluxed overnight. 1N HCl andethyl acetate were added to the reaction mixture, the organic phase wasseparated and dried (MgSO₄) and the solvent was removed to give thetitle compound GG.

1-(1H-Pyrazolo[3,4-b]pyridin-3-yl)-ethanone (HH)

H₂SO₄ (1 mL, 98%), H₂O (5 ml) and acetic acid (7.5 ml) was added to asolution of a crude reaction product mixture of2-(1H-pyrazolo[3,4-b]pyridine-3-carbonyl)-malonic acid dimethyl ester(GG). The mixture was heated at 120° C. for 3 h. The reaction mixturewas adjusted to pH 6.5 and ethyl acetate was used to extract theproduct. The product was purified by chromatography with 50%/50%hexanes/ethyl acetate to give the title compound (HH). H NMR (CDCl₃):2.80(s, 3H), 7.38(dd, 1H), 8.8.71(dd, 1H), 8.76(dd, 1H).

2-Bromo-1-(1H-pyrazolo[3,4-b]pyridin-3-yl)-ethanone (II)

To a solution of 1-(1H-pyrazolo[3,4-b]pyridin-3-yl)-ethanone (HH) (0.66g, 4.09 mol) in HBr (8 ml, 48%), bromine (0.654 g, 4.09 mmol) was addedand the reaction mixture was stirred at 65° C. for 2 h. A precipitatewas formed and filtered to give the HBr salt of the title compound (II)(0.94 g, 71% yield). H NMR (DMSO): 4.93(s, 2H), 7.45(dd, 1H), 8.56(dd,1H), 8.68(dd, 1H).

4-(1 H-Pyrazolo[3,4-b]pyridin-3-yl)-thiazol-2-ylamine (JJ)

An ethanol solution of2-Bromo-1-(1H-pyrazolo[3,4-b]pyridin-3-yl)-ethanone HBr salt (II) (0.94g, 2.92 mol) and thiourea (0.446 g, 5.84 mol) was stirred at roomtemperature overnight. A precipitate was formed and filtered to give thetitle compound (JJ). H NMR (DMSO): 7.03(s, bd 2H), 7.37(dd, 1H), 7.52(s,1H), 8.6(m, 2H).

Preparation ofN-(4-(1H-pyrazolo[3,4-b]pyridin-3-yl)thiazol-2-yl)-2-phenylacetamides

A suspension of 4-(1-H-pyrazolo[3,4-b]pyridin-3-yl)thiazol-2-ylamine(JJ) (0.065 g, 0.3 mmol), a phenylacetic acid optionally substitutedwith R (0.45 mmol), and Et₃N (0.091 g, 0.9 mmol) in 2 mL THF is heatedat 160° C. for 750 sec in microwave. The final product KK is purified bypreparative HPLC.

Example 5 Preparation ofN-(4-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)thiophen-2-yl)-2-(3-methsulfonamidophenyl)acetamide

5-(2-(trimethylsilyl)ethynyl)pyrimidin-4-amine (MM)

Trimethylsilylacetylene (3.4 mL) is added to a stirred suspension of LL(J. Heterocyclic Chem. 19: 1285, 1982; J. Org. Chem. 48: 1064, 1983;25.12 mmol), Pd(PPh₃)₂Cl₂ (1.7 g), CuI (0.95 g) in triethyamine (20 mL)and THF (50 mL) at room temperature and the solution is heated at 45° C.for 2 h. The solid is filtered off and the filtrate is concentrated togive a dark brown liquid. The crude product is purified by BiotageHorizon™ to afford title compound MM.

7H-pyrrolo[2,3-d]pyrimidine (NN)

A solution of NM (14.81 mmol) in THF (25 mL) is added to a stirredsuspension of potassium tert butoxide (2.5 g, 22.2 mmol) in THF (50 mL).The solution is refluxed for 3 h and then stirred at room temperaturefor 16 h. The solvent is evaporated and the residue is dissolved inwater (100 mL) and aqueous layer is extracted with ethyl acetate (5×50mL). Organic layer is dried and concentrated to give title compound NN.

5-iodo-7H-pyrrolo[2,3-d]pyrimidine (OO)

A 1M solution of iodine monochloride (5.26 mL, 5.26 mmol) is added to astirred solution of NN (2.63 mmol) in CH₂Cl₂ (25 mL) and the solution isstirred at room temperature for 16 h. The precipitated yellow solid isfiltered and suspended in ethyl acetate (50 mL) and washed withsaturated NaHCO₃ (25 mL). The organic layer is dried and concentrated togive title compound OO.

N-(4-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)thiophen-2-yl)-2-(3-methsulfonamidophenyl)acetamide(PP)

A mixture of 5-iodo-7H-pyrrolo[2,3-d]pyrimidine (OO) (1 mmol) and5-(2-(3-methsulfonamidophenyl)acetamido)thiophen-3-yl-3-boronic acid (E)(1.1 mmol), K₂CO₃ (3 mmol) and Pd(PPh₃)4 (5%) in the 3 mL dioxane and 1mL H₂O is heated in microwave to 150° C. for ten minutes. The organiclayer is separated and concentrated and the title compound PP ispurified by preparative HPLC.

Example 6 Preparation ofN-(4-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)thiophen-2-yl)-2-(3-methsulfonamidophenyl)acetamide

1H-Pyrazolo[3,4-d]pyrimidin-3-amine (RR)

4-aminopyrimidine-5-carbonitrile (QQ) (1 mmol) in 10 mL conc. HCl iscooled at 0° C., to which 1.2 equiv of NaNO₂ in 1 mL H₂O is addeddropwise and reaction mixture is stirred at 0° C. for 10 min. To thisreaction mixture, 5 equivalent of SnCl₂ in 5 mL conc. HCl is added andreaction mixture is stirred at room temperature overnight. 30 mL ofEtOAc is added to the reaction mixture and then washed with 1N NaOH. Theorganic layer is separated and dried. The solvent is removed to givetitle compound 1H-pyrazolo[3,4-d]pyrimidin-3-amine (RR).

3-iodo-1H-pyrazolo[3,4-d]pyrimidine (SS)

1H-pyrazolo[3,4-d]pyrimidin-3-amine (RR) (1 mmol) in 3 mL of sulfuricacid (16N) is cooled to 0° C., then 1.2 mmol of NaNO₂ in 1 mL of H₂O ofwater is added slowly at 0° C. The reaction mixture is stirred at 0° C.for 1 h. 4 mmol of potassium iodide in 3 mL H₂O is poured all at once.The mixture is warmed to room temperature over 1 h then brought to pH of7.8 using solid sodium carbonate and extracted with dichloromethane. Theorganic phase is washed with saturated solution of sodium thiosulfate,dried on MgSO₄, and finally evaporated under reduced pressure. The crudeproduct is purified on silica gel column chromatography to obtain thetitle compound (SS).

N-(4-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)thiophen-2-yl)-2-(3-methsulfonamidophenyl)acetamide(TT)

A mixture of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine (SS) (1 mmol) and5-(2-(3-methsulfonamidophenyl)acetamido)thiophen-3-yl-3-boronic acid (E)(1.1 mmol), K₂CO₃ (3 mmol) and Pd(PPh₃)₄ (5%) in the 3 mL dioxane and 1mL H₂O is heated in microwave to 150° C. for ten minutes. The organiclayer is separated and the title compoundN-(4-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)thiophen-2-yl)-2-(3-methsulfonamidophenyl)acetamide (TT) is purified by preparative HPLC.

Example 7 Preparation ofN-(5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-yl)-2-(3-methoxyphenyl)acetamide

5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-amine (VV)

A mixture of 7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (UU) (SeeNucleosides, Nucleotides & Nucleic Acids 20: 1823, 2001; 7 mmol) andthiosemicarbazide (2.34 g, 21 mmol) in trifluoroacetic acid (25 mL) isheated in a sealed tube at 100° C. for 2 hr. The brown solution iscooled to room temperature and poured into ice. The mixture is thenbasified with concentrated NH₄OH and the pale brown precipitate formedis filtered on a sintered glass filter. The solid is washed thoroughlywith water (3×50 mL) and ethyl acetate (3×50 mL) and dried under vacuumto afford 5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-amine(VV).

N-(5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-yl)-2-(3-methoxyphenyl)acetamide(WW)

A suspension of5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-amine (VV) (0.23mmol), 3-methoxyphenylacetic acid (0.038 g, 0.23 mmol), triethylamine(0.1 ML) and BtSO₂CH₃ (0.055 g, 0.28 mmol) in THF (3 mL) and DMF (0.3mL) is heated in a microwave oven for 20 min. The brown solution isadded to water (50 mL) and ethyl acetate (50 mL). Brine (10 ml) is addedto separate the layers. The organic phase is separated and washed withwater (2×50 mL). The organic layer is concentrated to give a solid,which is placed in a small Buchner funnel and washed with methanol (2×5mL) and ethyl acetate (2×5 mL). The brown solid collected is dried undervacuum to afford the desired productN-(5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-yl)-2-(3-methoxyphenyl)acetamide(WW).

Example 8 Preparation ofN-(5-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1,3,4-thiadiazol-2-yl)-2-(3-methoxyphenyl)acetamide

5-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1,3,4-thiadiazol-2-amine (YY)

A mixture of 1H-pyrazolo[3,4-d]pyrimidine-3-carbonitrile (XX) (SeeNucleosides, Nucleotides & Nucleic Acids 20: 1823, 2001; 7 mmol) andthiosemicarbazide (2.34 g, 21 mmol) in trifluoroacetic acid (25 mL) isheated in a sealed tube at 100° C. for 2 hr. The brown solution iscooled to room temperature and poured into ice. The mixture is thenbasified with concentrated NH₄OH and the pale brown precipitate formedis filtered on a sintered glass filter. The solid is washed thoroughlywith water (3×50 mL) and ethyl acetate (3×50 mL) and dried under vacuumto afford 5-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1,3,4-thiadiazol-2-amine(YY).

N-(5-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1,3,4-thiadiazol-2-yl)-2-(3-methoxyphenyl)acetamide(ZZ)

Compound ZZ is synthesized in a manner similar to compound WW usingamine YY instead to give the desired compound ZZ.

Example 9 Preparation of4-(1H-pyrazolo[3,4-b]pyridin-3-yl)-1H-pyrrole-2-carboxamides

(2-chloropyridin-3-yl)1-(5-(2,2,2-trichloroacetyl)-1H-pyrrol-3-yl)methanone(B′)

Aluminum(III) chloride (20 mmol) and compound A′ (Aldrich) are dissolvedin 10 mL dry CH₂Cl₂ at 0° C. 2,2,2-trichloro-1-(1H-pyrrol-2-yl)ethanone(10 mmol) is added dropwise and the reaction mixture is warmed to roomtemperature and stirred for 2 hours. The reaction mixture is poured intoice water and extracted with EtOAc. The organic layer is subsequentlywashed with saturated NaHCO₃ and brine. After drying and evaporation ofthe organic solvent, the mixture is passed through flash columnchromatography to give title compound B′ (540 mg, 15% yield).

Preparation of Compound C′

Compound B′ (60 mg) is dissolved into CH₃CN (2 mL), to which RNH₂ (1.2equivalent) is added, followed by triethyl amine (1.5 equivalent). Thereaction mixture is stirred at room temperature overnight. After thesolvent is evaporated, the reaction mixture is carried on to the nextstep without further purification.

Preparation of Compound D′

Compound C′ (reaction mixture) is dissolved in 2 mL EtOH, to which 3equivalent of hydrazine is added. The reaction mixture is heated in themicrowave at 170° C. for 10 minutes. The reaction mixture is directlyapplied on HPLC for purification to give title compound D′.

It will be appreciated that a variety of compounds can be preparedaccording to the general methods described above.

Example 10 Analytical Results

Table 2 below depicts exemplary LC mass spectral data (LC/MS), retentiontime (RT) and ¹H-NMR data (NMR) for certain compounds of the presentinvention, wherein compound numbers in Table 2 correspond to thecompounds depicted in Table 1 (empty cells indicate that the test wasnot performed): TABLE 2 Cmpd # LC/MS RT NMR 1 2 336.00 3.20 DMSO d-6:3.85(s, 2H), 7.28(m, 2H), 7.39(m, 4H), 7.68(s, 1H), 8,57(d, 1H),8.72(dd, 1H), 12.52(s, br, 1H), 13.83(s, br,1H) 3 366.10 3.23 DMSO d-6:3.77(s, 3H0, 3.80(s, 2H), 6.82(m, 1H), 6.93(m 2H), 7.3(m, 2H), 7.68(s,1H), 8.56(d, 1H), 8.7(dd, 1H)12.47(s, br, 1H), 13.75(s, br, 1H) 4 471.103.50 DMSO d-6: 1.03(t, 6H), 3.15(q, 4H, 3.65(s, 2H), 7.20(m, 2H), 7.5(m,1H), 7.6(m, 2H), 7.78(s, 1H), 8.48(dd, 1H), 8.72(dd, 1H) 5 429.10 2.79DMSO d-6: 2.93(s, 3H), 3.76(s, 2H), 6.99(d, 1H), 7.04(dd, 1H), 7.15(s,1H), 7.25(dd, 1H), 7.29(dd 1H), 7.64(s, 1H), 8.55(dd, 1H), 8.7(dd, 1H) 6446.00 2.30 DMSO-d6: 11.4(brS, 1H), 8.52(m, 2H), 7.57(s, 1H), 7.27(s,1H), 7.22(m, 1H), 6.72(m, 3H), 3.57(s, 2H), 3.17(s, 3H) 7 476.00 2.36CD3OD: 8.54(m, 2H), 7.51(d, 1H), 7.33(d, 1H), 7.29(dd, 1H), 7.23(t, 1H),6.91(m, 2H), 6.82(m, 1H), 4.01(t, 2H), 3.70(s, 1H), 3.34(m, 2H), 2.95(m,2H), 1.96(m, 2H), 1.81(m, 2H), 1.65(m, 1H), 1.49(m, 2H), 1.34(m, 2H) 8428.00 2.91 CD3OD: 8.52(m, 2H), 7.51(s, 1H), 7.14- 7.33(m, 6H), 3.74(s,2H), 2.96(s, 3H). 9 447.00 2.79 DMSO-d6: 13.98(s, 1H), 11.58(s, 1H),9.67(s, 1H), 8.68(d, 1H), 8.62(d, 1H), 7.83(s, 1H), 7.53(d, 1H),7.15-7.20(m, 3H), 3.77(s, 1H), 2.79(s, 3H). 10 442.00 3.02 CD3OD:8.57(d, 1H), 8.55(d, 1H), 7.80(s, 1H), 7.68(m, 2H), 7.60(t, 1H), 7.53(d,1H), 7.34(d, 1H), 7.30(dd, 1H), 3.88(s, 1H), 2.69(s, 6H) 11 411.00 2.32DMSO: 11.80 s (1H), 9.72 s (1H), 8.71 t (1H), 8.50 m (2H), 7.53 m (1H),7.49 m (1H), 7.30 t (1H), 7.20 m (2H), 7.10 m (2H), 4.45 d (2H), 2.97 s(3H) 12 286.00 1.71 MeOD: 8.72 d (1H), 8.60 d (1H), 7.60 s (1H), 7.42 s(1H), 7.34 m (1H), 4.18 m (1H), 3.59 m (2H), 1.26 d (3H) 13 348.00 2.64MeOD: 8.58 m (2H), 7.60 s (1H), 7.40 s (1H), 7.24 m (2H), 6.94 m (2H),6.80 d (1H),4.55 (2H), 3.78 s (3H) 14 402.10 2.60 MeOD: 8.54 m (2H),7.60 s (1H), 7.40 m (3H), 7.28 m (2H), 7.15 d (1H), 4.64 s (2H) 15362.10 2.59 MeOD: 8.60 m (2H), 7.60 s (1H), 7.38 s (1H), 7.29 dd (1H),6.87 m (2H), 6.76 m (1H), 5.91 s (2H), 4.47 s (2H) 16 335.90 2.71 MeOD:8.56 m (2H), 7.60 s (1H), 7.39 m (3H), 7.30 m (1H), 7.05 t (2H), 4.55 s(2H) 17 324.00 2.95 MeOD: 8.55 m (2H), 7.56 s (1H), 7.36 s (1H), 7.27 m(1H), 3.20 d (2H), 1.78 m (4H), 1.67 m (2H), 1.26 m (3H), 1.0 m (2H) 18336.00 2.70 MeOD: 8.55 m (2H), 7.60 s (1H), 7.40 s (1H), 7.34 m (1H),7.29 m (1H), 7.18 d (1H), 7.11 d (1H), 6.97 m (1H), 4.58 s (2H) 19476.00 1.70 CD3OD: 8.44(d, 1H), 8.25 (d, 1H), 7.97(s, 1H), 7.52(s, 1H),7.33(s, 1H), 7.23(t, 1H), 6.91(m, 2H), 6.81(dd,1H), 4.01(t, 2H), 3.68(s,2H), 3.36(m, 2H), 2.94(m, 2H), 1.93(m, 2H), 1.80(m, 2H), 1.68(brm, 1H),1.49(m, 2H), 1.36(m, 2H). 20 428.00 2.78 CD3OD: 8.44(d, 1H), 8.29(d,1H), 8.01(s, 1H), 7.31(m, 2H), 7.26(s, 1H), 7.17(d, 1H), 7.14(d, 1H),3.72(s, 2H), 2.96(s, 3H)

Example 11 ROCK Inhibition Assay

Compounds are screened for their ability to inhibit ROCK I (AA 6-553)activity using a standard coupled enzyme system (Fox et al. Protein Sci.7: 2249, 1998). Reactions are carried out in a solution containing 100mM HEPES (pH 7.5), 10 mM MgCl₂, 25 mM NaCl, 2 mM DTT and 1.5% DMSO.Final substrate concentrations in the assay are 45 μM ATP (SigmaChemicals, St Louis, Mo.) and 200 μM peptide (American Peptide,Sunnyvale, Calif.). Reactions are carried out at 30° C. and 45 nM ROCKI. Final concentrations of the components of the coupled enzyme systemare 2.5 mM phosphoenolpyruvate, 350 μM NADH, 30 μg/ml pyruvate kinaseand 10 μg/ml lactate dehydrogenase.

Compounds are screened for their ability to inhibit ROCK using astandard radioactive enzyme assay. Assays are carried out in a solutioncontaining 100 mM HEPES (pH 7.5), 10 mM MgCl₂, 25 mM NaCl, 2 mM DTT and1.5% DMSO. Final substrate concentrations in the assay are 13 μM [γ-³³P]ATP (25 mCi ³³P ATP/mmol ATP, Perkin Elmer, Cambridge, Mass./SigmaChemicals, St Louis, Mo.) and 27 μM Myelin Basic Protein (MBP). Finalenzyme concentration in the assay is 5 nM ROCK. Assays are carried outat room temperature. 1.5 μl of DMSO stock containing serial dilutions ofthe compound of the present invention (concentrations ranging from 10 μMto 2.6 nM) is placed in a 96 well plate. 50 μl of Solution 1 (100 mMHEPES (pH 7.5), 10 mM MgCl₂, 26 mM [γ-³³P] ATP) is added to the plate.The reaction is initiated by addition of 50 μl of Solution 2 (100 mMHEPES (pH 7.5), 10 mM MgCl₂, 4 mM DTT, 54 mM MBP and 10 nM ROCK). After2 hours the reaction is quenched with 50 μL of 30% trichloroacetic acid(TCA, Fisher) containing 9 mM ATP. Transfer of 140 μL of the quenchedreaction to a glass fiber filter plate (Corning, Cat. No. 3511) isfollowed by washing 3 times with 5% TCA. 50 μL of Optima Goldscintillation fluid (Perkin Elmer) is added and the plates are countedon a Top Count (Perkin Elmer). After removing mean background values forall of the data points the data is fit using Prism software to obtain aK_(i)(app).

Table 3 depicts enzyme inhibition data (K_(i)) for certain exemplarycompounds. Compound numbers in Table 3 correspond to those compoundsdepicted in Table 1.

In Table 3, “A” represents a K_(i) of less than 0.5 μM and “B”represents a K_(i) of between 0.5 and 5.0 μM. the term “Enzyme”indicates that an enzyme-linked assay was used; the term “³³P” indicatesthat a radioactive assay was used. TABLE 3 Cmpd # ROCK ³³P ROCK Enzyme 1A 2 A 3 A 4 A 5 A 6 A 7 A 8 A 9 A 10 A 11 A 12 B 13 A 14 A 15 A 16 A 17A 18 A 19 A 20 A

1. A compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein: wherein

Z₁, Z₂, Z₃ and Z₄ are each independently selected from N or CR¹, whereinat least one of Z₁, Z₂, or Z₄ is N; each R¹ is independently selectedfrom H, halogen, —CN, —NO₂, or —V_(m)R′; G is —NR²— or —CO—; Q¹ is —CO—,—SO₂—, —NR², —NR²CO—, —CONR²—, —SO₂NR²—, or is a bond; R² is —U_(n)R″;R³ is Q²-Ar¹, or when G is —NR², R² and Q¹-R³, taken together with thenitrogen atom, may form the cyclic group:

where s is 1 or 2, Z is CH or N; wherein each occurrence of Y isindependently —CO—, —CS—, —SO₂—, —O—, —S—, —NR⁵—, or —C(R⁵)₂—, and R⁵ isU_(n)R′; X₁ and X₂ are each independently selected from CR⁴ or N; eachoccurrence of R⁴ is independently selected from halogen, CN, NO₂, orV_(m)R; each occurrence of U or V is independently selected from anoptionally substituted C₁₋₆ alkylidene chain, wherein up to twomethylene units of the chain are optionally and independently replacedby —NR—, —S—, —O—, —CS—, —CO₂—, —OCO—, —CO—, —COCO—, —CONR—, —NRCO—,—NRCO₂—, —SO₂NR—, —NRSO₂—, —CONRNR—, —NRCONR—, —OCONR—, —NRNR—,—NRSO₂NR—, —SO—, —SO₂—, —PO—, —PO₂—, or —POR—; m and n are eachindependently 0 or 1; each occurrence of R is independently selectedfrom hydrogen or a C₁₋₆ aliphatic group, wherein said aliphatic group isoptionally substituted with up to five occurrences of J^(R); eachoccurrence of R′ is independently selected from hydrogen, a C₁₋₆aliphatic group, a 3-8-membered saturated, partially unsaturated, orfully unsaturated monocyclic ring having 0-3 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, or an 8-12 memberedsaturated, partially unsaturated, or fully unsaturated bicyclic ringsystem having 0-5 heteroatoms independently selected from nitrogen,oxygen, or sulfur, wherein said aliphatic group, monocyclic ring orbicyclic ring is optionally substituted with up to five occurrences ofJ^(R′); R″ is selected from hydrogen, a C₁₋₆ aliphatic group, a3-8-membered saturated, partially unsaturated, or fully unsaturatedmonocyclic ring having 0-3 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partiallyunsaturated, or fully unsaturated bicyclic ring system having 0-5heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein said aliphatic group, monocyclic ring or bicyclic ring isoptionally substituted with up to five occurrences of J^(R″); or twooccurrences of R, R′ and R″, in any combination thereof, are takentogether with the atom(s) to which they are bound to form a 3-12membered saturated, partially unsaturated, or fully unsaturatedmonocyclic or bicyclic ring having 0-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, wherein said monocyclic orbicyclic ring is optionally substituted with J^(R); each occurrence ofJ^(R), J^(R′) and J^(R″) is independently selected from halogen, L,-(L_(p))-R^(J), -(L_(p))-N(R^(J))₂, -(L_(p))-SR^(J), -(L_(p))-OR^(J),-(L_(p))-(C₃₋₁₀ cycloaliphatic), -(L_(p))-(C₆₋₁₀ aryl), -(L_(p))-(5-10membered heteroaryl), -(L_(p))-(5-10 membered heterocyclyl), oxo,C₁₋₄haloalkoxy, C₁₋₄haloalkyl, -(L_(p))-NO₂, -(L_(p))-CN, -(L_(p))-OH,-(L_(p))-CF₃, —CO₂R^(J), —CO₂H, —COR^(J), —COH, —OC(O)R^(J) or—NC(O)R^(J); or any two J^(R), J^(R′) or J^(R″) groups, on the samesubstituent or different substituents, together with the atom(s) towhich each J^(R), J^(R′) or J^(R″) group is bound, form a 5-7 memberedsaturated, unsaturated, or partially saturated ring; R^(J) is H or C₁₋₆aliphatic; or two R^(J) groups or an R^(J) group and an R, R′ or R″group, together with the atom to which they are attached, optionallyform a 3-6 membered cycloaliphatic or heterocyclyl, wherein saidaliphatic, cycloaliphatic or heterocyclyl is optionally substituted withR*, —OR*, —SR*, —NO₂, —CF₃, —CN, —CO₂R*, —COR*, OCOR* or NHCOR*, whereinR* is H or an unsubstituted C₁₋₆ aliphatic; L is a C₁₋₆ aliphaticwherein up to three methylene units are replaced by —NH—, —NR^(L)—, —O—,—S—, —CO₂—, —OC(O)—, —C(O)CO—, —C(O)—, —C(O)NH—, —C(O)NR⁶—, —C(═N—CN),—NHCO—, —NR^(L)CO—, —NHC(O)O—, —NR^(L)C(O)O—, —SO₂NH—, —SO₂NR^(L)—,—NHSO₂—, —NR^(L)SO₂—, —NHC(O)NH—, —NR^(L)C(O)NH—, —NHC(O)NR^(L)—,—NR^(L)C(O)NR^(L), —OC(O)NH—, —OC(O)NR^(L)—, —NHSO₂NH—, —NR^(L)SO₂NH—,—NHSO₂NR^(L)—, —NR^(L)SO₂NR^(L)—, —SO— or —SO₂—; R^(L) is selected fromC₁₋₆ aliphatic, C₃₋₁₀ cycloaliphatic, C₆₋₁₀ aryl, 5-10 memberedheteroaryl or 5-10 membered heterocyclyl; or two R^(L) groups, on thesame substituent or different substituents, together with the atom(s) towhich each R^(L) group is bound, form a 3-8 membered heterocyclyl; eachp is independently 0 or 1; Q² and Q³ are each independently selectedfrom a bond or a C₁₋₆ alkylidene chain, wherein up to two methyleneunits of the chain are each optionally and independently replaced by—NR′—, —S—, —O—, —CS—, —CO₂—, —OCO—, —CO—, —COCO—, —CONR′—, —NR′CO—,—NR′CO₂—, —SO₂NR′—, —NR′SO₂—, —CONR′NR′—, —NR′CONR′—, —OCONR′—,—NR′NR′—, —NR′SO₂NR′—, —SO—, —SO₂—, —PO—, —PO₂—, or —POR′—; and whereinany carbon atom in the one or more methylene units is optionallysubstituted with one or two occurrences of R⁶, wherein each occurrenceof R⁶ is independently halogen, —CN, —NO₂, or —U_(n)R′, or twooccurrences of R⁶, or R′ and R⁶, taken together with the atoms to whichthey are bound, form an optionally substituted 3-6-membered cycloalkyl,heterocyclyl, aryl or heteroaryl ring; and Ar¹ and Ar² are eachindependently selected from a C₁₋₆ aliphatic, a 3-8 membered saturated,partially unsaturated, or fully unsaturated monocyclic ring having 0-3heteroatoms independently selected from nitrogen, oxygen, or sulfur, oran 8-12 membered saturated, partially unsaturated, or fully unsaturatedbicyclic ring system having 0-5 heteroatoms independently selected fromnitrogen, oxygen, or sulfur; wherein Ar¹ and Ar² are each optionallysubstituted with 0-5 independent occurrences of TR⁷; wherein T is a bondor is a C₁-C₆ alkylidene chain wherein up to two methylene units of Tare optionally and independently replaced by —NR—, —S—, —O—, —CS—,—CO₂—, —OCO—, —CO—, —COCO—, —CONR—, —NRCO—, —NRCO₂—, —SO₂NR—, —NRSO₂—,—CONRNR—, —NRCONR—, —OCONR—, —NRNR—, —NRSO₂NR—, —SO—, —SO₂—, —PO—,—PO₂—, or —POR—; and each occurrence of R⁷ is independently selectedfrom —R′, halogen, —NO₂, —CN or ═O.
 2. The compound according to claim1, having one of formulae II-VII:

wherein R^(1Z1), R^(1Z2), R^(1Z3) and R^(1Z4) are each independentlyselected from H, halogen, —CN, —NO₂, or —V_(m)R′. 3-4. (canceled)
 5. Thecompound according to claim 1, wherein X₁ is CR⁴ and X₂ is N or CR⁴. 6.(canceled)
 7. The compound according to claim 1, wherein each R¹ isindependently selected from H, halogen or C₁₋₃ aliphatic. 8-13.(canceled)
 14. The compound according to claim 1, wherein Q¹ is —CO—,—SO₂—, —NR², —NR²CO—, —CONR²—, —SO₂NR².
 15. The compound according toclaim 14, wherein G is —NR² and Q¹ is —CO—, or G is —CO— and Q¹ is—NR²—.
 16. The compound according to claim 15, wherein R² is H, —C₁₋₄aliphatic, -cyclopropyl, (CH₂)₁₋₃OH or


17. (canceled)
 18. The compound according to claim 1, wherein R³ isQ²-Ar¹.
 19. The compound according to claim 18, wherein Q² is—(CHR⁶)_(q)—, —(CHR⁶)_(q)O—, —(CHR⁶)_(q)S—, —(CHR⁶)_(q)S(O)₂—,—(CHR⁶)_(q)S(O)—, —(CHR⁶)_(q)NR—, or —(CHR⁶)_(q)C(O)—, wherein q is 0,1, 2, or 3, and each R⁶ is R′, —N(R)(R′), —(CH₂)₁₋₄N(R)(R′),—(CH₂)₁₋₄C(CH₃)₂N(R)(R′), —(CH₂)₁₋₄CH(CH₃)N(R)(R′), —OR′, —(CH₂)₁₋₄OR′,—NR(CH₂)₁₋₄N(R)(R′), —NR(CH₂)₁₋₄SO₂R′, —NR(CH₂)₁₋₄COOR′, or—NR(CH₂)₁₋₄COR′, or two occurrences of R⁶, taken together with the atomsto which they are bound, form an optionally substituted 3-6-memberedsaturated, partially unsaturated, or fully unsaturated ring. 20-21.(canceled)
 22. The compound according to claim 18, wherein Ar¹ is a C₃₋₆aliphatic, a 5-8 membered saturated, partially unsaturated, or fullyunsaturated monocyclic ring having 0-3 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, or an 8-12 memberedsaturated, partially unsaturated, or fully unsaturated bicyclic ringsystem having 0-5 heteroatoms independently selected from nitrogen,oxygen, or sulfur; wherein Ar¹ is optionally substituted with 0-5independent occurrences of TR⁷.
 23. The compound according to claim 22,wherein Ar¹ is

wherein t is 0, 1, 2, 3, 4, or 5, and wherein any Ar¹ is bonded to Q²through any substitutable nitrogen or carbon atom, and wherein one ormore hydrogen atoms on any substitutable nitrogen or, carbon atom issubstituted with one or more independent occurrences of TR⁷. 24-26.(canceled)
 27. The compound according to claim 22, wherein TR⁷ isselected from —F, —Cl, —CN, —NH₂, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —OR^(x),—OCF₃, —NR^(x)SO₂R^(x), —NR^(x)SO₂N(R^(x))₂, —COOC(CH₃)₃, —OSO₂CH₃, —OH,—SO₂N(R^(x))₂, —SO₂N(R^(x))₂, —SO₂R^(x), -pyrollidinone, tetrahydrofuranor -D-(CH₂)_(p)—Y, wherein R^(x) is a H or a C₁₋₄ alkyl, D is —SO₂—,—SO₂NH—, —NHSO₂— or —O—, p is 0-3, and Y is selected from:

wherein R^(y) is H or C₁₋₃ alkyl, and wherein one or more carbon atomsof Y is optionally substituted with ═O. 28-31. (canceled)
 32. Thecompound according to claim 1, wherein said compound has a structuredepicted in Table
 1. 33. A composition comprising an effective amount ofcompound according to claim 1, and a pharmaceutically acceptablecarrier, adjuvant, or vehicle.
 34. The composition of claim 33,additionally comprising a therapeutic agent selected from achemotherapeutic or anti-proliferative agent, an anti-inflammatoryagent, an immunomodulatory or immunosuppressive agent, a neurotrophicfactor, an agent for treating cardiovascular disease, an agent fortreating destructive bone disorders, an agent for treating liverdisease, an anti-viral agent, an agent for treating blood disorders, anagent for treating diabetes, or an agent for treating immunodeficiencydisorders.
 35. A method of inhibiting ROCK kinase activity in abiological sample; which method contacting said biological sample with acompound of claim 1 or a composition comprising said compound.
 36. Amethod of treating or lessening the severity of a disease condition ordisorder selected from a proliferative disorder, a cardiac disorder, aneurodegenerative disorder, a psychotic disorder, an autoimmunedisorder, a condition associated with organ transplant, an inflammatorydisorder, an immunologically mediated disorder, a viral disease, or abone disorder, comprising the step of administering to said patient acompound according to claim 1 or a composition comprising said compound.37. The method of claim 36, comprising the additional step ofadministering to said patient an additional therapeutic agent selectedfrom a chemotherapeutic or anti-proliferative agent, ananti-inflammatory agent, an immunomodulatory or immunosuppressive agent,a neurotrophic factor, an anti-psychotic agent, an agent for treatingcardiovascular disease, an agent for treating destructive bonedisorders, an agent for treating liver disease, an anti-viral agent, anagent for treating blood disorders, an agent for treating diabetes, oran agent for treating immunodeficiency disorders, wherein: saidadditional therapeutic agent is appropriate for the disease beingtreated; and said additional therapeutic agent is administered togetherwith said composition as a single dosage form or separately from saidcomposition as part of a multiple dosage form. 38-40. (canceled)